C-to-G Transversion DNA Base Editors

ABSTRACT

Engineered transversion base editors that enable expanded amino acid modifications and methods of using the same. Described herein, for example, are fusion proteins containing cytidine deaminases (e.g. human or rat APOBECs, pmCDA1 or AID) or adenosine deaminases (e.g.  E. coli  TadAs) or a combination thereof, catalytically impaired CRISPR-Cas proteins (e.g. Cas9, CasX or Cas12 nucleases), linkers, nuclear localization signals (NLSs) and a human or  E. coli  uracil-n-glycosylase (UNG) and/or REV1 protein that enable the CRISPR-guided programmable introduction of C-to-G and G-to-C transversions in DNA. The UNG may be fused to the deaminase-Cas fusion or not, in which case endogenous UNG may be recruited using molecular machinery that is integrated into the deaminase-Cas fusion architecture, e.g. using peptide or RNA aptamers or scFVs, sdABs or Fabs.

CLAIM OF PRIORITY

This application claims the benefit of U.S. Patent Application Ser. No. 62/894,628 filed on Aug. 30, 2019; 62/910,912 filed on Oct. 4, 2019; 62/916,654 filed on Oct. 17, 2019; and 63/023,208, filed on May 11, 2020. The entire contents of the foregoing are hereby incorporated by reference.

FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

This invention was made with Government support under Grant No. HG009490 awarded by the National Institutes of Health and contract HR0011-17-2-0042 awarded by the Defense Advanced Research Projects Agency of the Department of Defense. The Government has certain rights in the invention.

TECHNICAL FIELD

Described herein are fusion proteins containing cytidine deaminases (e.g. human or rat APOBECs, pmCDA1 or AID) or adenosine deaminases (e.g. E. coli TadAs) or a combination thereof, catalytically impaired CRISPR-Cas proteins (e.g. Cas9, CasX or Cas12 nucleases), linkers, nuclear localization signals (NLSs) and a human or E. coli uracil-n-glycosylase (UNG) and/or REV1 protein that enable the CRISPR-guided programmable introduction of C-to-G and G-to-C transversions in DNA. The UNG may be fused to the deaminase-Cas fusion or not, in which case endogenous UNG may be recruited using molecular machinery that is integrated into the deaminase-Cas fusion architecture, e.g. using peptide or RNA aptamers or scFVs, sdABs or Fabs.

BACKGROUND

DNA base editors represent a new class of genome editing tools that enable the programmable installation of single or multiple base substitutions. Current generations of cytosine base editors (CBE) and adenine base editors (ABE) allow for the targeted deamination of cytosines and adenines that get exposed on ssDNA by RNA-guided CRISPR-Cas proteins¹⁻⁴. The majority of disease-associated genetic perturbations known to date are point mutations, also known as single nucleotide variants (SNVs). Current iterations of CBEs and ABEs can target disease-relevant transition mutations and revert them to the original genotype, e.g. correcting G-to-A (C-to-T) mutations using ABE. However, a relevant fraction of disease-associated SNVs represent C-to-G and G-to-C substitutions that cannot be targeted using current BEs.

SUMMARY

Described herein are CRISPR-guided C-to-G transversion base editors (CGBE) that enable the installation of cytosine-to-guanine and guanine-to-cytosine base edits in the ssDNA bubble generated by RNA-guided fusion proteins that contain adenine (e.g. E. coli TadA) and/or cytosine (e.g. rat APOBEC1) deaminases as well as CRISPR-Cas proteins (e.g. S. pyogenes Cas9) and/or REV1 or UNG proteins that are directly fused and/or recruited to the deaminase-Cas fusion protein. CGBE comprises a programmable DNA-binding domain (e.g. catalytically impaired dead or nicking Cas9) fused to a cytosine and/or adenosine deaminase. The adenosine deaminase can be a wild type (WT) or mutant E. coli TadA or previously described engineered TadA variants in the form of monomers, homodimers or heterodimers thereof, to decrease RNA editing activity while still preserving DNA editing activity (SECURE or RRE variants, Grünewald et al, NBT 2019—in press). The cytidine deaminase can be, e.g. rat APOBEC1, A3A, AID or pmCDA1, or previously described engineered variants of these deaminases (e.g. rAPOBEC1 with mutations from SECURE-BE3) with reduced RNA editing activity and preserved DNA editing capabilities⁵⁻⁹. In some embodiments, CGBE comprises one or more uracil-N-glycosylases (UNGs) fused to the N and/or C-terminus of the CBE or ABE fusion protein without uracil-N-glycosylase inhibitors (UGIs) and potentially with fused REV1 proteins. In some embodiments, CGBE comprise a linker between the adenosine or cytidine deaminase and the programmable DNA binding domain as well as between the deaminase domain and the UNG or the DNA binding domain and the UNG. In some embodiments the TadA domain can be monomeric, homodimeric or heterodimeric and contain all combinations of wild type (WT) E. coli TadA, or mutant variants of TadA).

Thus, provided herein are C-to-G transversion base editors (CGBEs) comprising a cytidine deaminase, a programmable DNA binding domain, and further comprising one or more nuclear localization sequences (NLS), and optionally one or more human or E. coli or other uracil-n-glycosylases (UNGs) or SMUG1, preferably wherein the CGBE does not comprise a uracil-N-glycosylase inhibitors (UGI).

In some embodiments, the cytidine deaminase comprises an active cytidine deaminase domain, preferably a monomeric domain, from a wild type and/or engineered rat APOBEC1 (rAPOBEC1), human APOBEC3A, human APOBEC3G, human AID, pmCDA1 (e.g., shown in Tables A and B) or variations thereof bearing mutations that reduce RNA or DNA off-target editing while retaining efficient DNA base editing.

In some embodiments, the cytidine deaminase comprises one or more mutations corresponding to mutations in rAPOBEC1, human APOBEC3A, human APOBEC3G, human AID or pmCDA1 or in any homologue or orthologue thereof (optionally those in Tables A and B).

In some embodiments, the cytidine deaminase is a rAPOBEC1 or any one of its ortho- or paralogues listed in Tables A or B, comprises one or more mutations that decrease RNA editing activity while preserving DNA editing activity, wherein the mutations are at amino acid positions that correspond to residues R33, P29, K34, E181, and/or L182 of rAPOBEC1 (SEQ ID NO:67) or to W90Y, R126E, R132E, W90Y+R126E (double mutant), R126E+R132E (double mutant), W90Y+R132E (double mutant), W90Y+R126E+R132E (triple mutant) (see, e.g., Ref. 16).

In some embodiments, the one or more mutations comprises a mutation at amino acid position that correspond to: (1) residue R33 of WT rAPOBEC1 or evoAPOBEC1; or (2) residue R13 in evoFERNY-APOBEC1; or (3) residue R12 in FERNY-APOBEC1.

In some embodiments, the mutation at amino acid position that correspond to residue R33 is a R33A substitution mutation.

In some embodiments, the CGBE comprises N- or C-terminal fusions of one or more human or E. coli UNG or SMUG1 or other orthologues of UNG or SMUG1 (e.g. as shown in Table J).

In some embodiments, the one or more UNGs are E. coli UNGs.

In some embodiments, the UNG(s) is absent, e.g., to minimize indel formation and reduce the size/length of the editor (e.g. miniCGBE1).

In some embodiments, the cytidine deaminase is a wildtype or engineered rAPOBEC1 (or any one of its ortho- or paralogues listed in Tables A or B) and the cytidine deaminase bears one or more mutations at positions: P29F, P29T, R33A, K34A, R33A+K34A (double mutant), E181Q and/or L182A of rAPOBEC1 (SEQ ID NO:67).

In some embodiments, the CGBE further includes one or more mutations at its cytidine deaminase rAPOBEC1 (or any one of its ortho- or paralogues listed in Tables A or B) residues corresponding to E24, V25; R118, Y120, H121, R126; W224-K229; P168-1186; L173+L180; R15, R16, R17, to K15-17 &A15-17; Deletion E181-L210; P190+P191; Deletion L210-K229 (C-terminal); and/or Deletion S2-L14 (N-terminal) of SEQ ID NO:67.

In some embodiments, the CGBE does not comprise one or more UNGs and/or the CGBE further comprises translesion polymerase REV1 (SEQ ID NO: 200) on either the N- or C-terminus or on both. In some embodiments, the CGBE comprises one or more UNGs and the tvBE further comprises a translesion polymerase REV1 (SEQ ID NO: 200). In some embodiments, the translesion polymerase REV1 (SEQ ID NO: 200) is fused to either the N- or C-terminus or both.

In some embodiments, the CGBE includes a linker between the cytosine deaminase monomer and/or between the cytosine deaminase monomer or single-chain dimers and the programmable DNA binding domain.

Exemplary Constructs Include:

1. CGBE1:

bpNLS-E.coliUNG-LINKER-rAPOBEC1(R33A)-LINKER-SpCas9(D10A)-bpNLS

2. miniCGBE1:

bpNLS-rAPOBEC1(R33A)-LINKER-SpCas9(D10A)-bpNLS

In some embodiments, the programmable DNA binding domain is selected from the group consisting of an engineered C2H2 zinc-finger, a transcription activator effector-like effector (TALE), and a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) Cas RNA-guided nuclease (RGNs) and variants thereof.

The CGBE of any one of claims 1-15, wherein the CRISPR RGN is a ssDNA nickase or a catalytically inactive CRISPR Cas RNA-guided nuclease (e.g., a Cas9 or Cas12a that has ssDNA nickase activity or is catalytically inactive); in some embodiments, the Cas RGN is from SpCas9-NG or VRQR-Cas9.

Also provided herein are base editing systems comprising:

(i) a CGBE as described herein, wherein the programmable DNA binding domain is a CRISPR Cas RGN or a variant thereof; and (ii) at least one guide RNA compatible with the base editor comprising a spacer sequence that directs the base editor to a target sequence, preferably wherein the target sequence comprises a cytosine at position 4-8, 5-7, or position 6 (with 1 being the most PAM-distal position).

Also provided herein are isolated nucleic acids encoding a CGBE as described herein, vectors comprising the isolated nucleic acids, and isolated host cells, preferably mammalian host cells (but also plant, bacterial, etc), comprising the nucleic acids or the vectors described herein. In some embodiments, the isolated host cell expresses the CGBE of any one of claims 1-17.

Additionally provided herein are methods for generating a cytosine-to-guanine and guanine-to-cytosine alteration in a nucleic acid, the method comprising contacting the nucleic acid with the CGBE of any one of claims 1-17, or the base editing system of claim 18.

In some embodiments, the CGBE achieves at least 15%, at least 20%, at least 25%, at least 30%, at least 35%, at least 40%, at least 45%, least 50%, at least 55%, at least 60%, or at least 63% C-to-G conversions in a target sequence.

In some embodiments, the target sequence is a sequence within or adjacent to one of the genes in Table E1 or Table E2.

Also provided herein are methods for generating a cytosine-to-guanine and guanine-to-cytosine alteration in a selected nucleotide of a target region of a nucleic acid. The methods include contacting the nucleic acid with:

(i) a C-to-G transversion base editor (CGBE) comprising an adenosine deaminase, e.g., a wild type and/or engineered (e.g. ABEs 0.1, 0.2, 1.1, 1.2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 2.10, 2.11, 2.12, 3.1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7, 3.8, 4.1, 4.2, 4.3, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 5.10, 5.11, 5.12, 5.13, 5.14, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 7.10, ABEmax) E. coli TadA monomer, or variations of homo- or heterodimers thereof, bearing one or more mutations in either or both monomers that decrease RNA editing activity while preserving DNA editing activity, wherein the mutations are at amino acid positions that correspond to residues of E. coli TadA as listed in Table H, a programmable DNA binding domain comprising a ssDNA nickase or a catalytically inactive CRISPR Cas RNA-guided nuclease; and

(ii) at least one guide RNA compatible with the base editor and comprising a spacer that directs the base editor to the target sequence, preferably wherein the target sequence comprises a cytosine at position 4-8, 5-7, or position 6 (with 1 being the most PAM-distal position).

In some embodiments, the cytosine-to-guanine or guanine-to-cytosine alteration is listed in Table D.

Also provided herein are compositions comprising a CGBE or base editing systems as described herein, optionally including one or more ribonucleoprotein (RNP) complexes.

Additionally provided herein are the CGBE or base editing systems described herein, for use in generating a cytosine-to-guanine and guanine-to-cytosine alteration in a cell, wherein the alteration corrects a specific disease-related mutation provided in Tables E1 and E2.

In some embodiments, the CGBE does not comprise a UNG, and the CGBE recruits endogenous UNG with the help of a peptide aptamer fused to the CGBE.

In some embodiments, the CGBE does not comprise a UNG, and CGBE recruits endogenous UNG with the help of RNA aptamers fused to the gRNA.

In some embodiments, the CGBE does not comprise a UNG, and the CGBE recruits endogenous UNG with the help of a Fab, scFV or sdAb elements fused to the CGBE.

In some embodiments, the CGBE does not comprise a UNG, and wherein the CGBE recruits endogenous REV1 translesion polymerase.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Methods and materials are described herein for use in the present invention; other, suitable methods and materials known in the art can also be used. The materials, methods, and examples are illustrative only and not intended to be limiting. All publications, patent applications, patents, sequences, database entries, and other references mentioned herein are incorporated by reference in their entirety. In case of conflict, the present specification, including definitions, will control.

Other features and advantages of the invention will be apparent from the following detailed description and figures, and from the claims.

DESCRIPTION OF DRAWINGS

FIGS. 1A-D. C-to-G transversion at position C6 in the FANCF site 1 spacer as an on-target byproduct of ABEmax and miniABEmax treatment in human HEK293T cells. FIG. 1A. Efficient DNA on-target A-to-G editing of the adenine in position 4 of the spacer (with 1 being the most PAM-distal position) by ABEmax and two miniABEmax variants compared to a nCas9-only negative control. 1B. C-to-G editing of the DNA cytosine in position 6 of the FANCF site 1 spacer in all ABE variants tested in the same experiment as shown in FIG. 1A. 1C. C-to-T editing of the DNA cytosine in position 6 of the FANCF site 1 spacer in all ABE variants tested in the same experiment as shown in FIG. 1A. 1D. C-to-A editing of the DNA cytosine in position 6 of the FANCF site 1 spacer in all ABE variants tested in the same experiment as shown in a. All data generated from independent quadruplicate experiments (n=4).

FIGS. 2A-2C. C-to-G transversion at position C6 is the predominant on-target byproduct on three genomic sites in human HEK293T cells treated with ABEmax and miniABEmax. 2A. C-to-G editing of the DNA cytosine in position 6 of the HEK site 2, ABE site 7, and FANCF site 1 spacer in all ABE variants tested with FANCF site 1 exhibiting the highest editing efficiencies as shown in FIGS. 1A-D. 2B. C-to-T editing of C6 was seen only at FANCF site 1. 2C. C-to-A editing in position 6 was only seen at consistently high levels at around 1-5% at FANCF site 1.

FIG. 3 . Potential mechanism of action explaining C-to-G editing byproducts induced by ABE treatment in human HEK293T cells—part I. Schematic of an ABEmax protein inducing parallel targeted A-to-I deamination in the target ssDNA bubble as well as potentially inducing byproduct C-to-U deamination on position 6 of the spacer.

FIG. 4 . Potential mechanism of action explaining C-to-G editing byproducts induced by ABE treatment in human HEK293T cells—part II. Schematic of uracil excision by UNG after the byproduct C-to-U deamination on position 6 was induced by ABE, leading to an abasic site at position 6 of the spacer. Downstream activity of mismatch repair (MMR) pathways and of the translesion polymerase REV1 as well as secondary deamination of adenines in C-to-A byproducts could potentially explain the higher proportion of C-to-G outcomes in position 6.

FIG. 5 . Schematic drawing of approach to increase C-to-G product. Leveraging downstream processing of abasic sites by e.g. MMR and REV1, we propose using a CBE fusion protein containing a cytidine deaminase to enhance C-to-U deamination compared to ABE. In contrast to conventional CBE architectures, we propose to exchange the UGIs for a single or multiple UNG proteins to further increase the creation of abasic sites, thereby increasing the input for potential MMR and REV1 processing that may eventually lead to improved C-to-G editing yield.

FIG. 6 . Schematic drawing of a C-to-G transversion base editor (CGBE) architecture. An N-terminal deaminase domain, e.g. rAPOBEC1, FERNY-APOBEC1, evoFERNY-APOBEC1, evoAPOBEC1, AID, A3A, eA3A, pmCDA1, A3G or an E. coli TadA mutant was fused to a catalytically impaired DNA binding protein, e.g. dCas9 or Cas9 nickase (D10A). An E. coli or human UNG protein was fused to the C-terminus.

FIG. 7 . Schematic drawing of a C-to-G transversion base editor (CGBE) architecture that can show reduced indel byproduct frequency by fusing bacteriophage Mu Gam protein. The depicted fusion proteins showed a highly similar composition as the construct in FIG. 6 with the exception of the N-terminal (or C-terminal) fusion of the bacteriophage Mu Gam protein to reduce indel fractions, i.e. also in combination with the use of catalytically inactive Cas9 (dCas9).

FIG. 8 . Schematic drawing of a C-to-G transversion base editor (CGBE) architecture with a fusion of the translesion polymerase REV1. In this construct, the anatomy of the initial CGBE (FIG. 6 ) was altered by exchanging UNG for REV1 on the C- or N-terminus.

FIG. 9 . Schematic drawing of a C-to-G transversion base editor (CGBE) architecture with a fusion of both UNG and the translesion polymerase REV1. In this construct, the anatomy of the initial CGBE (FIG. 6 ) was altered by adding REV1 on the C- or N-terminus, leading to a CGBE variant that contains both UNG and REV1 as a direct fusion.

FIG. 10 . Schematic drawing showing a construct where the anatomy of the initial CGBE (FIG. 6 ) was altered by fusing a peptide aptamer to the C- or N-terminus in order to recruit endogenous UNG instead of directly fusing UNG to the deaminase-Cas9 fusion protein.

FIG. 11 . Schematic drawing showing a construct where the anatomy of the initial CGBE (FIG. 6 ) was altered by fusing a scFV, Fab or sdAb to the C- or N-terminus in order to recruit endogenous UNG instead of directly fusing UNG to the deaminase-Cas9 fusion protein.

FIG. 12 . Schematic drawing showing a construct where the anatomy of the initial CGBE (FIG. 6 ) was altered by encoding an RNA aptamer directly in the gRNA in order to recruit endogenous UNG instead of directly fusing UNG to the deaminase-Cas9 fusion protein.

FIG. 13 . Engineering of a C-to-G base editor. Bar plots showing on-target DNA base editing frequencies with various base editor architectures using seven gRNAs targeting genomic sites in HEK293T cells. N and C indicate amino-terminal and carboxy-terminal ends, respectively, of the various base editors. Gray overlay bars at top represent deletions at each editing window. Target cytosines are highlighted. Editing frequencies of three independent replicates (n=3) at each base are displayed side-by-side. Percentage values below specific cytosine bases indicate the average C-to-G editing observed (values below 3% not reported). Numbering on the bottom indicates position of the base in the protospacer with 1 being the most PAM-distal base. Arrowheads indicate cytosines showing C-to-G edits. Arrows point at examples of C-to-G edits.

FIG. 14 . On-target activities of nCas9 controls, ABE variants, and more CBE variants tested for C-to-G editing in HEK293T cells. Bar plots showing the on-target DNA base editing frequencies induced by nCas9 negative controls, ABE and ABE variants, and other CBE variants with seven gRNAs in HEK293T cells. N and C indicate amino-terminal and carboxy-terminal ends, respectively, of the various base editors. Gray overlay bars at top represent deletions at each editing window. Editing frequencies of three independent replicates (n=3) at each base are displayed side-by-side. Percentage values below specific cytosine bases indicate the average C-to-G editing observed (values below 3% not reported). Numbering on the bottom indicates position of the base in the protospacer with 1 being the most PAM-distal base. Arrowheads indicate cytosines showing C-to-G edits.

FIGS. 15A-B. Indel frequencies of nCas9 controls, ABE variants, and CBE variants tested for C-to-G editing in HEK293T cells. a,b, Dot plots representing percentage of alleles that contain an insertion or deletion across the entire protospacer from experiments with various base editor architectures reported in FIG. 14 (15 a) or FIGS. 13 and 14 (15 b). Single dots represent individual replicates.

FIG. 16 . On-target activities of non-APOBEC1 CBE variants tested for C-to-G editing in HEK293T cells. Bar plots showing the on-target DNA base editing frequencies induced by non-APOBEC1 CBEs and their variants with h/eUNG with seven gRNAs in HEK293T cells. N and C indicate amino-terminal and carboxy-terminal ends, respectively, of the various base editors. Gray overlay bars at top represent deletions at each editing window. Editing frequencies of three independent replicates (n=3) at each base are displayed side-by-side. Percentage values below specific cytosine bases indicate the average C-to-G editing observed (values below 3% not reported). Numbering on the bottom indicates position of the base in the protospacer with 1 being the most PAM-distal base. Arrowheads indicate cytosines showing C-to-G edits.

FIG. 17 . Indel frequencies of non-APOBEC1 CBE variants tested for C-to-G editing in HEK293T cells. Dot plots representing percentage of alleles that contain an insertion or deletion across the entire protospacer from experiments with non-APOBEC1 CBE variants reported in FIG. 16 . Single dots represent individual replicates.

FIGS. 18A-B. Additional characterization of CGBE1 on-target editing activities in HEK293T cells. A,B, Bar plots showing the on-target DNA base editing frequencies induced by BE4max(R33A) and CGBE1 using 12 gRNAs with a C at position 6 (C6-sites; 18A) and 6 gRNAs with a C at position 4, 5, 7, or 8 (non-C6-sites; 18B) in HEK293T cells. N and C indicate amino-terminal and carboxy-terminal ends, respectively, of the various base editors. Gray overlay bars at top represent deletions at each editing window. Editing frequencies of three independent replicates (n=3) at each base are displayed side-by-side. Percentage values below specific cytosine bases indicate the average C-to-G editing observed (values below 3% not reported). Numbering on the bottom indicates position of the base in the protospacer with 1 being the most PAM-distal base. Arrowheads indicate cytosines showing C-to-G edits.

FIG. 19 . Aggregated distribution of editing and indel frequencies across protospacer of BE4max(R33A) and CGBE1 in HEK293T cells. Dot and box plots representing the combined distribution of C-to-G, C-to-T, C-to-A, and indel frequencies (labeled) across the entire protospacer from experiments performed with BE4max(R33A) and CGBE1 using 25 guides. Boxes span the interquartile range (IQR; first to third quartiles), horizontal lines indicate the median (second quartile), and whiskers extend to ±1.5×IQR. Single dots represent individual replicates. The graphs were derived from the data shown in FIGS. 13 and 18A-B.

FIGS. 20A-B. On-target activities of nCas9 controls and CGBE1-related variants with more gRNAs in HEK293T cells. A,B, Bar plots showing the on-target DNA base editing frequencies of nCas9 controls and CGBE1-related variants using 12 gRNAs with a C at position 6 (C6-sites; 20A) and 6 gRNAs with a C at position 4, 5, 7, or 8 (non-C6-sites; 20B) in HEK293T cells. N and C indicate amino-terminal and carboxy-terminal ends, respectively, of the various base editors. Gray overlay bars at top represent deletions at each editing window. Editing frequencies of three independent replicates (n=3) at each base are displayed side-by-side. Percentage values below specific cytosine bases indicate the average C-to-G editing observed (values below 3% not reported). Numbering on the bottom indicates position of the base in the protospacer with 1 being the most PAM-distal base. Arrowheads indicate cytosines showing C-to-G edits.

FIG. 21 . Indel frequencies of CGBE1 and CGBE1-related variants with more gRNAs in HEK293T cells. Dot plots representing percentage of alleles that contain an insertion or deletion across the entire protospacer from experiments with CGBE1-related variants reported in FIGS. 18A-B and 20A-B. Single dots represent individual replicates.

FIGS. 22A-B. Comparison of CGBE1 and miniCGBE1 on-target editing activities with 25 gRNAs in HEK293T cells. A,B, Bar plots showing the on-target DNA base editing frequencies of CGBE1 and miniCGBE1 using 19 gRNAs with a C at position 6 (C6-sites; 22A) and 6 gRNAs with a C at position 4, 5, 7, or 8 (non-C6-sites; 22B) in HEK293T cells. N and C indicate amino-terminal and carboxy-terminal ends, respectively, of the various base editors. Gray overlay bars at top represent deletions at each editing window. Editing frequencies of three independent replicates (n=4) at each base are displayed side-by-side. Percentage values below specific cytosine bases indicate the average C-to-G editing observed (values below 3% not reported). Numbering on the bottom indicates position of the base in the protospacer with 1 being the most PAM-distal base. Arrowheads indicate cytosines showing C-to-G edits.

FIGS. 23A-B. On-target activities of nCas9 control with 25 gRNAs in HEK293T cells. A,B, Bar plots showing the on-target DNA base editing frequencies observed with expression of a nCas9 negative control using 19 gRNAs with a C at position 6 (C6-sites; 23A) and 6 gRNAs with a C at position 4, 5, 7, or 8 (non-C6-sites; 23B) in HEK293T cells. N and C indicate amino-terminal and carboxy-terminal ends, respectively, of the various base editors. Gray overlay bars at top represent deletions at each editing window. Editing frequencies of three independent replicates (n=4) at each base are displayed side-by-side. Percentage values below specific cytosine bases indicate the average C-to-G editing observed (values below 3% not reported). Numbering on the bottom indicates position of the base in the protospacer with 1 being the most PAM-distal base. Arrowheads indicate cytosines showing C-to-G edits in respective CGBE experiments.

FIG. 24 . Indel frequencies of CGBE1 and miniCGBE1 variants with 25 gRNAs in HEK293T cells. Dot plots representing percentage of alleles that contain an insertion or deletion across the entire protospacer from experiments with CGBE1 and miniCGBE1 reported in FIG. 22 and control experiments reported in FIG. 23 . Single dots represent individual replicates.

FIG. 25 . Additional comparison of CGBE1 and miniCGBE1 on-target editing activities with 23 non-C6 gRNAs in HEK293T cells. Bar plots showing the on-target DNA base editing frequencies induced by nCas9 control, BE4max, BE4max(R33A), CGBE1, and miniCGBE1 with 23 gRNAs for sites with a C at position 4, 5, 7, or 8 (non-C6 sites) in HEK293T cells. N and C indicate amino-terminal and carboxy-terminal ends, respectively, of the various base editors. Gray overlay bars at top represent deletions at each editing window. Editing frequencies of three independent replicates (n=3) at each base are displayed side-by-side. Percentage values below specific cytosine bases indicate the average C-to-G editing observed (values below 3% not reported). Numbering on the bottom indicates position of the base in the protospacer with 1 being the most PAM-distal base. Arrowheads indicate cytosines showing C-to-G edits.

FIG. 26 . Indel frequencies of CGBE1 and miniCGBE1 variants with 23 non-C6 gRNAs in HEK293T cells. Dot plots representing percentage of alleles that contain an insertion or deletion across the entire protospacer from experiments with BE4max, BE4max(R33A), CGBE1 and miniCGBE1 reported in FIG. 25 . Single dots represent individual replicates.

FIGS. 27A-B. Aggregated distribution of C-to-G editing frequencies across protospacer of CGBE1 and miniCGBE1 in HEK293T cells. A,B, Dot and box plots representing the aggregate distribution of C-to-G (yellow) editing frequencies across the entire protospacer from experiments performed with CGBE1 (27A) and miniCGBE1 (27B) with all 48 tested gRNAs. Boxes span the interquartile range (IQR; first to third quartiles), horizontal lines indicate the median (second quartile), and whiskers extend to ±1.5×IQR. Single dots represent individual replicates. The graphs were derived from the data shown in FIGS. 22A-B and 25.

FIG. 28 . Off-target DNA editing activities of CGBE1 and miniCGBE1 in HEK293T cells. Bar plots showing the off-target DNA base editing frequencies induced by nCas9 control, BE4max, BE4max(R33A), CGBE1, and miniCGBE1 using HEK site 2, HEK site 3, HEK site 4, EMX1 site 1, and FANCF site 1 gRNAs in HEK293T cells. N and C indicate amino-terminal and carboxy-terminal ends, respectively, of the various base editors. Gray overlay bars at top represent deletions at each editing window. Editing frequencies of three independent replicates (n=3) at each base are displayed side-by-side. Percentage values below specific cytosine bases indicate the average C-to-D (D=A/T/G) editing observed (values below 1% not reported). Numbering on the bottom indicates position of the base in the protospacer with 1 being the most PAM-distal base. Arrowheads indicate cytosines showing C-to-G edits.

FIG. 29 . Indel frequencies of CGBE1 and miniCGBE1 variants for DNA off-targets in HEK293T cells. Dot plots representing percentage of alleles that contain an insertion or deletion across the entire protospacer from experiments with BE4max, BE4max(R33A), CGBE1 and miniCGBE1 reported in FIG. 28 . Single dots represent individual replicates.

FIG. 30 . On-target DNA editing activities of NG and VRQR variants of CGBE1 and miniCGBE1 in HEK293T cells. Bar plots showing the on-target DNA base editing frequencies induced by NG and VRQR variants of nCas9, CGBE1, and miniCGBE1 using 6 gRNAs that target AT-rich genomic loci with PAMs that are compatible with SpCas9-NG (NGT) and SpCas9-VRQR (NGAG) variants in HEK293T cells. N and C indicate amino-terminal and carboxy-terminal ends, respectively, of the various base editors. Gray overlay bars at top represent deletions at each editing window. Editing frequencies of three independent replicates (n=4) at each base are displayed side-by-side. Percentage values below specific cytosine bases indicate the average C-to-G editing observed (values below 3% not reported). Numbering on the bottom indicates position of the base in the protospacer with 1 being the most PAM-distal base. Arrowheads indicate cytosines showing C-to-G edits.

FIG. 31 . Indel frequencies of NG and VRQR variants of CGBE1 and miniCGBE1 variants in HEK293T cells. Dot plots representing percentage of alleles that contain an insertion or deletion across the entire protospacer from experiments with NG and VRQR variants of CGBE1 and miniCGBE1 reported in FIG. 30 . Single dots represent individual replicates.

FIG. 32 . Potential mechanism of prime editing system. Schematic of prime editing (PE) used to install a C-to-G substitution. PE fusion protein consists of an SpCas9-H840A nickase fused to an engineered Moloney murine leukemia virus reverse transcriptase (MMLV-RT). The prime editing guide RNA (pegRNA) consists of a standard targetable SpCas9 gRNA that also harbors a 3′ extension containing a primer binding site (PBS) and a reverse transcription template (RTT) that encodes the desired edit. PE2 system encompasses the prime editor fusion protein and a pegRNA. PE3 system additionally includes a nicking gRNA (ngRNA).

FIGS. 33A-B. Testing PE2 and PE3 in multiple human cell lines. A,B, Bar and dot plots representing the on-target DNA prime editing and indel frequencies of PE2 and PE3 targeting FANCF site 1 for G-to-T prime editing (33A) and HEK site 3 for PE-induced CTT insertion (33B) in 4 cell lines. Single dots represent individual replicates. Error bars represent standard deviation.

FIG. 34 . Comparing the editing activities of CGBEs and PEs in multiple human cell lines. Bar plots showing the average on-target DNA C-to-G base or prime editing frequencies induced by CGBE1, miniCGBE1, PE2, or PE3 on four genomic target loci. Each site in each cell line was tested with four independent replicates in HEK293T cells and three independent replicates in K562, U205, and HeLa cells. Single dots represent individual replicates. A two-tailed Student's t-test with p-values adjusted for multiple testing was used to calculate the shown p-values. Error bars represent standard deviations.

FIG. 35 . Testing pegRNAs and nicking gRNAs with wild-type SpCas9 in HEK293T cells. Bar and dot plots representing the frequency of alleles with indels (%) induced by pegRNAs and nicking gRNAs used in the experiments in FIGS. 33 and 34 (and FANCF site 1+21 ngRNA control) with wild-type SpCas9 in HEK293T. pegRNAs/ngRNAs designed by Anzalone et al. and by us are separated by the dashed line. Single dots represent individual replicates. Error bars represent standard deviations. ND, not done.

FIG. 36 . Additional comparisons of CGBE1, miniCGBE1, PE2, and PE3 on-target editing activities in HEK293T, K562, U2OS, and HeLa cells. Bar plots showing the on-target DNA editing frequencies induced by nCas9 controls, CGBE1, miniCGBE1, PE2, and PE3 with four gRNAs (CGBEs), four pegRNAs (PE2), or 4 pegRNA/nicking gRNA combinations (PE3), designed to install a C-to-G substitution at the same cytosine at four genomic loci in four cell lines. Gray overlay bars at top represent deletions at each site. Editing frequencies of four independent replicates (n=4) for HEK293T cells or three independent replicates (n=3) for K562, U2OS, and HeLa cells at each base are displayed side-by-side. Percentage values below cytosine bases reflect the average C-to-G editing observed (values below 3% not reported). Numbering on the bottom indicates position of the base with 1 being the most PAM-distal base for base editors, or the first nucleotide 3′ of the pegRNA/Cas9-induced nick for prime editors. Arrowheads indicate cytosines showing C-to-G edits.

FIG. 37 . Indel frequencies of CGBE1, miniCGBE1, PE2, and PE3 in HEK293T, K562, U2OS, and HeLa cells. Dot plots representing percentage of alleles that contain an insertion or deletion across the entire protospacer from experiments with CGBE1, miniCGBE1, PE2, and PE3 reported in FIGS. 34 and 36 . Single dots represent individual replicates.

DETAILED DESCRIPTION

ABEs install A-to-G substitutions in DNA while CBEs allow for the introduction of C-to-T mutations. However, both these types of mutations represent transitions and the extensive subset of disease-associated transversion mutations—e.g. C-to-G mutations-cannot be directly targeted with neither CBEs nor ABEs.

We sought to engineer a C-to-G transversion base editor (CGBE) that enables the programmable installation of C-to-G and G-to-C mutations. Based on our finding that ABE proteins that do not comprise UGIs can reproducibly induce C-to-G editing at position 6 of the spacer (with 1 being the most PAM-distal position) at mutliple genomic sites (FIGS. 1 and 2 ; Grunewald et al, Nature Biotechnology 2019), we hypothesized that we could engineer a base editing construct that might allow for higher C-to-G yield. We engineered CGBEs comprised of cytidine deaminases or adenosine deaminases or both (e.g. as in dual-deaminase architecture of bifunctional adenine and cytosine base editors, BACE) fused to DNA binding proteins (e.g. dCas9 or nickase Cas9) as well as to UNG or REV1 proteins or a combination thereof. We hypothesized that using a cytidine deaminase will increase C-to-U deamination rates at C6 or neighboring cytosines at the target ssDNA bubble, and fusing base excision repair (BER) protein UNG or translesion polymerase REV1 (without fusing a UGI) might enable increased formation of an abasic site at position 6 of the genomic target site. Downstream processing of the abasic site via MMR or translesion synthesis could subsequently yield higher C-to-G product (FIG. 3-5 ). Described herein are a number of different fusion protein architectures involving the abovementioned domains and proteins. Some embodiments use dCas9 and/or bacteriophage Mu Gam (FIG. 6-9 ; Komor et al, Sci Adv 2017) to reduce insertion/deletion (indel) byproducts, thereby further increasing relative C-to-G product yield and purity. In some embodiments, the methods include recruiting endogenous UNG to the programmable base editing target site with the use of peptide aptamers fused to CGBEs (delta UNG), RNA aptamers integrated into the gRNA or CGBE (delta UNG) fusion proteins harboring scFVs, sdABs or Fabs to recruit endogenous UNG (FIG. 10-12 ).

Thus, described herein are variants of base editor fusion proteins that enable the programmable introduction of transversion base edits, specifically C-to-G and G-to-C. A table of potentially actionable codon and amino acid changes are shown in Table D and a list of potential disease targets (using Cas proteins compatible with NGG, NG, and NGA-PAMs) is shown in Tables E1-E3.

Exemplary Cytidine Deaminase Domains Used for CGBE

In some embodiments, the cytidine deaminase is pmCDA1 (sea lamprey) or APOBEC1 from rat, or from a different species (Table A), e.g., a different mammalian species such as H. sapiens. The APOBEC, AICDA (AID) and CDA1 family members have high sequence homology and represent potential candidates for CGBE architectures (Table B)^(2,15-18).

Specifically, reduced RNA editing variants of rAPOBEC1, enhanced human A3A, and human AID are candidates for inclusion into CGBE architectures.

In some embodiments, CGBE described herein can be a wild-type BE4max or SECURE-BE4max-R33A as well as eA3A variants with truncated UGIs and additional N- or C-terminal fusion of a human or E. coli UNG.

In some embodiments, the cytidine deaminases in Anc-BE4max, evoAPOBEC1-BE4max (SEQ ID 205), FERNY-BE4max, evoFERNY-BE4max (SEQ ID 204), CDA1-BE4max, and evoCDA1-BE4max may be used in a BE4max architecture with truncated UGIs and optionally also have UNGs (human or E. coli, N- or C-terminal) added. In other embodiments, the SECURE-CBE R33 and/or K34 residue changes may be introduced in evoAPOBEC1.

In some embodiments, R13 and/or K14 residue changes are introduced in FERNY and evoFERNY-APOBEC1 (these residue changes are embedded in the same amino acid sequence motif as R33 and K34 in WT rat APOBEC1 that was used in BE3, BE4, and BE4max). These modifications (single or double residue change) can greatly reduce RNA off-target editing and enhance on-target C-to-G editing. All of the APOBEC1-based CBEs described herein can used with or without the proposed mutations in the context of a C-to-G transversion base editor.

The cytidine deaminase domain need not include an entire full protein, but can be a variant as described herein that has changes or truncations that do not abolish the cytidine deaminase activity.

Exemplary Adenosine Deaminase Domains Used for CGBE

In some embodiments, the adenosine deaminase is TadA from E. coli, or an orthologue from a different prokaryote, e.g. S. aureus, or a homologue from the eukaryotic domain, such as yeast TAD1/2 or a mammalian species such as human (e.g. ADAT2; Table C). The tRNA-specific adenosine deaminase family members have high sequence homology and many of these orthologues may be compatible with one or more of the amino acid substitutions in E. coli TadA expected to cause an RRE phenotype and would be desirable in a CGBE architecture.

The wild type sequence of wild type E. coli TadA, available in uniprot at P68398, is as follows:

(SEQ ID NO: 1) MSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGWNRPI GRHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSR IGRVVFGARDAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLSD FFRMRRQEIKAQKKAQSSTD.

The engineered E. coli TadA sequence present in ABE7.10 and ABEmax is as follows:

SEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIG LHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRI GRVVFGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALLCYF FRMPRQVFNAQKKAQSSTD.

In the most commonly used ABEs (ABE7.10 and ABEmax), these two proteins were fused using a 32 amino acid linker (bolded in sequence below), forming a heterodimer, the sequence of which is as follows:

MSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGWNRPI GRHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSR IGRVVFGARDAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLSD FFRMRRQEIKAQKKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSS GGSSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNR AIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIH SRIGRVVFGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALL CYFFRMPRQVFNAQKKAQSSTD.

Other exemplary sequences are shown in Table C. These tRNA-specific adenosine deaminase orthologues and homologues also represent candidates for inclusion of the mutations previously described at analogous positions in these proteins.

In some embodiments, the base editors included catalytically dead adenine deaminase variants, e.g. E59A. (Gaudelli et al, 2017, PMID: 29160308) as part of a heterodimer.

The adenine deaminase domain need not include an entire full protein, but can be a variant as described herein that has changes or truncations that do not abolish the adenine deaminase activity.

Uracil DNA Dlycosylase (UNG)

Cellular molecular pathways are in complete homeostasis within healthy cells. Especially, DNA repair pathways are balanced in ways that potentially mutagenic lesions are repaired at the optimal level. In mammalian cells, there is continuous generation of deamination mutations and repair of deamination reactions occurring in the background. Impairments in this process can lead to disruption of this homeostasis. On the deamination side, aberrant overexpression of deaminases that can induce spontaneous deamination at DNA and RNA levels has been shown to be responsible for inducing different cancers.^(10,11) On the other hand, expression levels of DNA glycosylases—a family of enzymes responsible for repairing the deaminated bases via the base excision repair (BER) pathway—are also crucial. DNA glycosylases carry out their activity by removing the lesions and creating abasic sites. Overexpression of uracil DNA glycosylase (UNG) has been shown to confer chemotherapy resistance in certain cancers.¹² Moreover, overexpression of uracil glycosylase inhibitor (UGI), a component of CBEs, is potentially responsible for the observed levels of toxicity and genome-wide Cas9-independent DNA off-target effects that can be induced by CBEs. In the light of these aforementioned independent observations, it is clear that one needs to control and optimize the expression levels of the exogeneous base editor constructs in order to minimize the potential unwanted side-effects to the target cells and preserve the homeostasis.

In some embodiments of the C-to-G transversion base editors (CGBEs) described herein, Uracil-DNA glycosylase (UNG) is a critical component that carries out the generation of abasic sites after cytosines are deaminated to uracil.

Exemplary UNG/SMUG Sequences for Inclusion in CGBE

In some embodiments, the CGBE fusion proteins described herein include a functional UNG or Single-Strand-Selective Monofunctional Uracil-DNA Glycosylase 1 (SMUG1) domain. Table J provides a list of UNG and SMUG1 orthologues.

Recruiting Endogenous UNG to Target and Edit Genetic Loci

While overexpression of engineered constructs is the first and main strategy to edit genomic loci, it has been well established that overexpression of exogeneous proteins can have unwanted and fatal consequences. In the context of base editors specifically, it has been demonstrated that overexpression of base editors can induce hundreds to thousands of off-target single nucleotide variations (SNVs) on DNA and RNA.^(6,7,13,14) All in all, there is great need to temporally and spatially control the expression levels of base editors in target cells. To this end, recruiting the endogenous cellular machinery to carry out the enzymatic reactions of interest, instead of exogenously providing a protein in excess, is a prominent bypass to minimize exogeneous components that need to be overexpressed.

It is possible that exogeneous overexpression of human or bacterial UNG may alter the repair pathway balance towards more efficient abasic site generation genome-wide. While more research is warranted to elucidate the impact of such UNG overexpression in mammalian cells, bypassing the need for overexpression of an immunogenic (in the case of E. coli UNG) protein and preserving the natural endogenous expression levels of UNG would be advantageous. To this end, we are proposing to utilize three alternative methods/constructs with the aim of recruiting the endogenous UNG to the target site of deamination.

Section 1: Peptide Aptamer Mediated Recruiting of UNG to the Target Site

Peptide aptamers are small amino acid sequences that can be designed and selected against virtually any given protein of interest. Peptide aptamers can have dissociation constants similar to naturally found antibodies. Owing to their small size, ease of production, high specificity, higher stability and solubility, peptide aptamers represent a significant alternative to the antibodies. Starting from an initial randomized library of peptides, peptide aptamers can be selected and further optimized via various methods in vitro and in vivo.

Fusing an engineered peptide aptamer against human UNG into our CGBE constructs would allow us to recruit endogenous UNG bypassing the need to overexpress the protein exogenously. (FIG. 10 )

Also, various peptide aptamers can be engineered from scratch against human UNG by methods including but not limited to yeast-two-hybrid systems in vivo, and phage-display in vitro systems. Candidate peptide aptamers displaying strong affinity against human UNG will be sequenced and the identified DNA and amino acid sequences will be employed as fusion partners in our next generation CGBE constructs. Optimal conformation of the peptide aptamer fusion will be determined empirically by cloning it into different sites in our constructs with different linkers.

Section 2: RNA Aptamer Mediated Recruiting of UNG to the Target Site

RNA aptamers are short stretches (80-120 nucleotides) of RNA molecules with strong and selective affinity against the target proteins of interest. Candidate RNA aptamers can be chemically synthesized as randomized libraries and several rounds of in vitro and in vivo selections can be applied. Employing the method called Systematic Evolution of Ligands by EXponential enrichment (SELEX), a number of candidate RNA aptamer molecules can be identified against one's target protein of interest.

As an example, the fusion of MS2 aptamers to CRISPR gRNAs is a widely used and well-known example of such a strategy. In this strategy, MS2 RNA aptamers are fused to the ends of gRNA constructs, thereby enabling specific recruitment of MS2 bacteriophage coat protein fused target proteins. Therefore, we propose that fusing an already engineered RNA aptamer against human UNG, if any exists, into the gRNA component of our CGBE constructs would allow us to recruit endogenous UNG bypassing the need to overexpress exogenously. (FIG. 12 )

Also, various RNA aptamers against human UNG can be engineered by strategies including but not limited to the available in vitro and in vivo SELEX strategies in the literature. Candidate RNA aptamers displaying strong affinity against human UNG will be sequenced and identified RNA sequences will be employed as gRNA fusion partners in our next generation CGBE constructs. Optimal conformation of the RNA aptamer fusion will be determined empirically by cloning it into different sites in our gRNA constructs with different linkers.

Section 3: Fab, scFV, or sdAb Mediated Recruiting of UNG to the Target Site

Antibodies are naturally expressed immunological proteins comprised of two light and two heavy chain proteins expressed from different genes. They are selected against specific parts (epitopes) of specific target proteins (antigens) in immune cells. Therefore, they can selectively bind to target antigens with high affinities. Antibodies are large molecules (˜150 kDa) consisting of a constant region (Fc) and antigen binding regions (Fab) with number of disulfide bonds in between chains. Therefore, it is not practical to generate a single peptide fusion protein fused with a large intact multimeric antibody and one's protein of interest.

However, getting rid of the Fc portion and using a single Fab portion of an antibody is a smaller (˜50 kDa) and more viable option to have than having a UNG fusion partner. Important to note is that the Fab portion still has constant regions of heavy and light chains that can be further resected while retaining the antigen specific binding affinity. This approach produces a shorter fragment (˜25 kdA) called single-chain variable fragment (scFv) that is linked with each other via short peptide linker. scFv consists of variable domains of heavy and light chains. Taking one step further and separating variable domains of heavy and light chains and producing a single chain (thus single variable domain) antibody fragment is called single-domain antibodies (sdAb) or nanobodies. This is the smallest of all antibody fragments (˜12-15 kDa) around 110 amino acids in length.

Given these premises, fusing an Fab, scFv or sdAb raised against human UNG target protein to our CGBE constructs in different conformations would be a viable option to recruit the endogenous human UNG to the target loci.

Also, various new Fabs, scFvs and sdAbs against human UNG can be generated by methods including but not limited to generating a mouse hybridoma clone, then converting full IgG (or IgM) into a scFv, Fab or sdAb; generating an immunized phage display scFv, Fab or sdAb mouse library, then using human UNG to screen the library; screening a premade scFv, Fab or sdAb antibody phage display library; generating synthetic libraries by altering the variable domains of antibodies via introducing random oligonucleotides, then screening against human UNG.

Candidate Fabs, scFvs or sdAbs displaying strong affinity against human UNG will be sequenced and the identified DNA and amino acid sequences will be employed as fusion partners in our next generation CGBE constructs. Optimal conformation of the fusion partners will be determined empirically by cloning it into different sites in our constructs with different linkers.

Programmable DNA Binding Domain

In some embodiments, the base editors include programmable DNA binding domains such as engineered C2H2 zinc-fingers, transcription activator effector-like effectors (TALEs), and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) Cas RNA-guided nucleases (RGNs) and their variants, including ssDNA nickases (nCas9) or their analogs and catalytically inactive dead Cas9 (dCas9) and its analogs (e.g., as shown in Table F), and any engineered protospacer-adjacent motif (PAM) or high-fidelity variants (e.g., as shown in Table G). A programmable DNA binding domain is one that can be engineered to bind to a selected target sequence.

CRISPR-Cas Nucleases

Although herein we refer to Cas9, in general any Cas9-like nickase could be used (including the related Cpf1/Cas12a enzyme classes), unless specifically indicated. These orthologs, and mutants and variants thereof as known in the art, can be used in any of the fusion proteins described herein. See, e.g., WO 2017/040348 (which describes variants of SaCas9 and SpCas 9 with increased specificity) and WO 2016/141224 (which describes variants of SaCas9 and SpCas 9 with altered PAM specificity).

The Cas9 nuclease from S. pyogenes (hereafter simply Cas9) can be guided via simple base pair complementarity between 17-20 nucleotides of an engineered guide RNA (gRNA), e.g., a single guide RNA or crRNA/tracrRNA pair, and the complementary strand of a target genomic DNA sequence of interest that lies next to a protospacer adjacent motif (PAM), e.g., a PAM matching the sequence NGG or NAG (Shen et al., Cell Res (2013); Dicarlo et al., Nucleic Acids Res (2013); Jiang et al., Nat Biotechnol 31, 233-239 (2013); Jinek et al., Elife 2, e00471 (2013); Hwang et al., Nat Biotechnol 31, 227-229 (2013); Cong et al., Science 339, 819-823 (2013); Mali et al., Science 339, 823-826 (2013c); Cho et al., Nat Biotechnol 31, 230-232 (2013); Jinek et al., Science 337, 816-821 (2012)). The engineered CRISPR from Prevotella and Francisella 1 (Cpf1, also known as Cas12a) nuclease can also be used, e.g., as described in Zetsche et al., Cell 163, 759-771 (2015); Schunder et al., Int J Med Microbiol 303, 51-60 (2013); Makarova et al., Nat Rev Microbiol 13, 722-736 (2015); Fagerlund et al., Genome Biol 16, 251 (2015). Unlike SpCas9, Cpf1/Cas12a requires only a single 42-nt crRNA, which has 23 nt at its 3′ end that are complementary to the protospacer of the target DNA sequence (Zetsche et al., 2015). Furthermore, whereas SpCas9 recognizes an NGG PAM sequence that is 3′ of the protospacer, AsCpf1 and LbCp1 recognize TTTN PAMs that are found 5′ of the protospacer (Id.).

In some embodiments, the present system utilizes a wild type or variant Cas9 protein from S. pyogenes or Staphylococcus aureus, or a wild type or variant Cpf1 protein from Acidaminococcus sp. BV3L6 or Lachnospiraceae bacterium ND2006 either as encoded in bacteria or codon-optimized for expression in mammalian cells and/or modified in its PAM recognition specificity and/or its genome-wide specificity. A number of variants have been described; see, e.g., WO 2016/141224, PCT/US2016/049147, Kleinstiver et al., Nat Biotechnol. 2016 August; 34(8):869-74; Tsai and Joung, Nat Rev Genet. 2016 May; 17(5):300-12; Kleinstiver et al., Nature. 2016 Jan. 28; 529(7587):490-5; Shmakov et al., Mol Cell. 2015 Nov. 5; 60(3):385-97; Kleinstiver et al., Nat Biotechnol. 2015 December; 33(12):1293-1298; Dahlman et al., Nat Biotechnol. 2015 November; 33(11):1159-61; Kleinstiver et al., Nature. 2015 Jul. 23; 523(7561):481-5; Wyvekens et al., Hum Gene Ther. 2015 July; 26(7):425-31; Hwang et al., Methods Mol Biol. 2015; 1311:317-34; Osborn et al., Hum Gene Ther. 2015 February; 26(2):114-26; Konermann et al., Nature. 2015 Jan. 29; 517(7536):583-8; Fu et al., Methods Enzymol. 2014; 546:21-45; and Tsai et al., Nat Biotechnol. 2014 June; 32(6):569-76, inter alia. Concerning rAPOBEC1 itself, a number of variants have been described, e.g. Chen et al, RNA. 2010 May; 16(5):1040-52; Chester et al, EMBO J. 2003 Aug. 1; 22(15):3971-82.: Teng et al, J Lipid Res. 1999 April; 40(4):623-35.; Navaratnam et al, Cell. 1995 Apr. 21; 81(2):187-95.; MacGinnitie et al, J Biol Chem. 1995 Jun. 16; 270(24):14768-75.; Yamanaka et al, J Biol Chem. 1994 Aug. 26; 269(34):21725-34. The guide RNA is expressed or present in the cell together with the Cas9 or Cpf1. Either the guide RNA or the nuclease, or both, can be expressed transiently or stably in the cell or introduced as a purified protein or nucleic acid.

In some embodiments, the Cas9 also includes one of the following mutations, which reduce nuclease activity of the Cas9; e.g., for SpCas9, mutations at D10A or H840A (which creates a single-strand nickase).

In some embodiments, the SpCas9 variants also include mutations at one of each of the two sets of the following amino acid positions, which together destroy the nuclease activity of the Cas9: D10, E762, D839, H983, or D986 and H840 or N863, e.g., D10A/D10N and H840A/H840N/H840Y, to render the nuclease portion of the protein catalytically inactive; substitutions at these positions could be alanine (as they are in Nishimasu al., Cell 156, 935-949 (2014)), or other residues, e.g., glutamine, asparagine, tyrosine, serine, or aspartate, e.g., E762Q, H983N, H983Y, D986N, N863D, N863S, or N863H (see WO 2014/152432).

In some embodiments, the Cas9 is fused to one or more SV40 or bipartite (bp) nuclear localization sequences (NLSs) protein sequences; an exemplary (bp)NLS sequence is as follows: (KRTADGSEFES)PKKKRKV (SEQ ID NO: 204). Typically, the NLSs are at the N- and C-termini of an ABEmax fusion protein, but can also be positioned at the N- or C-terminus in other ABEs, or between the DNA binding domain and the deaminase domain. Linkers as known in the art can be used to separate domains.

TAL Effector Repeat Arrays

Transcription activator like effectors (TALEs) of plant pathogenic bacteria in the genus Xanthomonas play important roles in disease, or trigger defense, by binding host DNA and activating effector-specific host genes. Specificity depends on an effector-variable number of imperfect, typically ˜33-35 amino acid repeats. Polymorphisms are present primarily at repeat positions 12 and 13, which are referred to herein as the repeat variable-diresidue (RVD). The RVDs of TAL effectors correspond to the nucleotides in their target sites in a direct, linear fashion, one RVD to one nucleotide, with some degeneracy and no apparent context dependence. In some embodiments, the polymorphic region that grants nucleotide specificity may be expressed as a triresidue or triplet.

Each DNA binding repeat can include a RVD that determines recognition of a base pair in the target DNA sequence, wherein each DNA binding repeat is responsible for recognizing one base pair in the target DNA sequence. In some embodiments, the RVD can comprise one or more of: HA for recognizing C; ND for recognizing C; HI for recognizing C; HN for recognizing G; NA for recognizing G; SN for recognizing G or A; YG for recognizing T; and NK for recognizing G, and one or more of: HD for recognizing C; NG for recognizing T; NI for recognizing A; NN for recognizing G or A; NS for recognizing A or C or G or T; N* for recognizing C or T, wherein * represents a gap in the second position of the RVD; HG for recognizing T; H* for recognizing T, wherein * represents a gap in the second position of the RVD; and IG for recognizing T.

TALE proteins may be useful in research and biotechnology as targeted chimeric nucleases that can facilitate homologous recombination in genome engineering (e.g., to add or enhance traits useful for biofuels or biorenewables in plants). These proteins also may be useful as, for example, transcription factors, and especially for therapeutic applications requiring a very high level of specificity such as therapeutics against pathogens (e.g., viruses) as non-limiting examples.

Methods for generating engineered TALE arrays are known in the art, see, e.g., the fast ligation-based automatable solid-phase high-throughput (FLASH) system described in U.S. Ser. No. 61/610,212, and Reyon et al., Nature Biotechnology 30,460-465 (2012); as well as the methods described in Bogdanove & Voytas, Science 333, 1843-1846 (2011); Bogdanove et al., Curr Opin Plant Biol 13, 394-401 (2010); Scholze & Boch, J. Curr Opin Microbiol (2011); Boch et al., Science 326, 1509-1512 (2009); Moscou & Bogdanove, Science 326, 1501 (2009); Miller et al., Nat Biotechnol 29, 143-148 (2011); Morbitzer et al., T. Proc Natl Acad Sci USA 107, 21617-21622 (2010); Morbitzer et al., Nucleic Acids Res 39, 5790-5799 (2011); Zhang et al., Nat Biotechnol 29, 149-153 (2011); Geissler et al., PLoS ONE 6, e19509 (2011); Weber et al., PLoS ONE 6, e19722 (2011); Christian et al., Genetics 186, 757-761 (2010); Li et al., Nucleic Acids Res 39, 359-372 (2011); Mahfouz et al., Proc Natl Acad Sci USA 108, 2623-2628 (2011); Mussolino et al., Nucleic Acids Res (2011); Li et al., Nucleic Acids Res 39, 6315-6325 (2011); Cermak et al., Nucleic Acids Res 39, e82 (2011); Wood et al., Science 333, 307 (2011); Hockemeye et al. Nat Biotechnol 29, 731-734 (2011); Tesson et al., Nat Biotechnol 29, 695-696 (2011); Sander et al., Nat Biotechnol 29, 697-698 (2011); Huang et al., Nat Biotechnol 29, 699-700 (2011); and Zhang et al., Nat Biotechnol 29, 149-153 (2011); all of which are incorporated herein by reference in their entirety.

Zinc Fingers

Zinc finger (ZF) proteins are DNA-binding proteins that contain one or more zinc fingers, independently folded zinc-containing mini-domains, the structure of which is well known in the art and defined in, for example, Miller et al., 1985, EMBO J., 4:1609; Berg, 1988, Proc. Natl. Acad. Sci. USA, 85:99; Lee et al., 1989, Science. 245:635; and Klug, 1993, Gene, 135:83. Crystal structures of the zinc finger protein Zif268 and its variants bound to DNA show a semi-conserved pattern of interactions, in which typically three amino acids from the alpha-helix of the zinc finger contact three adjacent base pairs or a “subsite” in the DNA (Pavletich et al., 1991, Science, 252:809; Elrod-Erickson et al., 1998, Structure, 6:451). Thus, the crystal structure of Zif268 suggested that zinc finger DNA-binding domains might function in a modular manner with a one-to-one interaction between a zinc finger and a three-base-pair “subsite” in the DNA sequence. In naturally occurring zinc finger transcription factors, multiple zinc fingers are typically linked together in a tandem array to achieve sequence-specific recognition of a contiguous DNA sequence (Klug, 1993, Gene 135:83).

Multiple studies have shown that it is possible to artificially engineer the DNA binding characteristics of individual zinc fingers by randomizing the amino acids at the alpha-helical positions involved in DNA binding and using selection methodologies such as phage display to identify desired variants capable of binding to DNA target sites of interest (Rebar et al., 1994, Science, 263:671; Choo et al., 1994 Proc. Natl. Acad. Sci. USA, 91:11163; Jamieson et al., 1994, Biochemistry 33:5689; Wu et al., 1995 Proc. Natl. Acad. Sci. USA, 92: 344). Such recombinant zinc finger proteins can be fused to functional domains, such as transcriptional activators, transcriptional repressors, methylation domains, and nucleases to regulate gene expression, alter DNA methylation, and introduce targeted alterations into genomes of model organisms, plants, and human cells (Carroll, 2008, Gene Ther., 15:1463-68; Cathomen, 2008, Mol. Ther., 16:1200-07; Wu et al., 2007, Cell. Mol. Life Sci., 64:2933-44).

One existing method for engineering zinc finger arrays, known as “modular assembly,” advocates the simple joining together of pre-selected zinc finger modules into arrays (Segal et al., 2003, Biochemistry, 42:2137-48; Beerli et al., 2002, Nat. Biotechnol., 20:135-141; Mandell et al., 2006, Nucleic Acids Res., 34:W516-523; Carroll et al., 2006, Nat. Protoc. 1:1329-41; Liu et al., 2002, J. Biol. Chem., 277:3850-56; Bae et al., 2003, Nat. Biotechnol., 21:275-280; Wright et al., 2006, Nat. Protoc., 1:1637-52). Although straightforward enough to be practiced by any researcher, recent reports have demonstrated a high failure rate for this method, particularly in the context of zinc finger nucleases (Ramirez et al., 2008, Nat. Methods, 5:374-375; Kim et al., 2009, Genome Res. 19:1279-88), a limitation that typically necessitates the construction and cell-based testing of very large numbers of zinc finger proteins for any given target gene (Kim et al., 2009, Genome Res. 19:1279-88).

Combinatorial selection-based methods that identify zinc finger arrays from randomized libraries have been shown to have higher success rates than modular assembly (Maeder et al., 2008, Mol. Cell, 31:294-301; Joung et al., 2010, Nat. Methods, 7:91-92; Isalan et al., 2001, Nat. Biotechnol., 19:656-660). In preferred embodiments, the zinc finger arrays are described in, or are generated as described in, WO 2011/017293 and WO 2004/099366. Additional suitable zinc finger DBDs are described in U.S. Pat. Nos. 6,511,808, 6,013,453, 6,007,988, and 6,503,717 and U.S. patent application 2002/0160940.

Variants

In some embodiments, the components of the fusion proteins are at least 80%, e.g., at least 85%, 90%, 95%, 97%, or 99% identical to the amino acid sequence of a exemplary sequence (e.g., as provided herein), e.g., have differences at up to 1%, 2%, 5%, 10%, 15%, or 20% of the residues of the exemplary sequence replaced, e.g., with conservative mutations, e.g., including or in addition to the mutations described herein. Optionally the differences can include truncations or deletions. In preferred embodiments, the variant retains a desired activity of the parent, e.g., deaminase activity, and/or the ability to interact with a guide RNA and/or target DNA, optionally with improved specificity or altered substrate specificity.

To determine the percent identity of two nucleic acid sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second amino acid or nucleic acid sequence for optimal alignment and non-homologous sequences can be disregarded for comparison purposes). The length of a reference sequence aligned for comparison purposes is at least 80% of the length of the reference sequence, and in some embodiments is at least 90% or 100%. The nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position (as used herein nucleic acid “identity” is equivalent to nucleic acid “homology”). The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which need to be introduced for optimal alignment of the two sequences. Percent identity between two polypeptides or nucleic acid sequences is determined in various ways that are within the skill in the art, for instance, using publicly available computer software such as Smith Waterman Alignment (Smith, T. F. and M. S. Waterman (1981) J Mol Biol 147:195-7); “BestFit” (Smith and Waterman, Advances in Applied Mathematics, 482-489 (1981)) as incorporated into GeneMatcher Plus™, Schwarz and Dayhof (1979) Atlas of Protein Sequence and Structure, Dayhof, M. O., Ed, pp 353-358; BLAST program (Basic Local Alignment Search Tool; (Altschul, S. F., W. Gish, et al. (1990) J Mol Biol 215: 403-10), BLAST-2, BLAST-P, BLAST-N, BLAST-X, WU-BLAST-2, ALIGN, ALIGN-2, CLUSTAL, or Megalign (DNASTAR) software. In addition, those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the length of the sequences being compared. In general, for proteins or nucleic acids, the length of comparison can be any length, up to and including full length (e.g., 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, or 100%). For purposes of the present compositions and methods, at least 80% of the full length of the sequence is aligned.

For purposes of the present disclosure, the comparison of sequences and determination of percent identity between two sequences can be accomplished using a Blosum 62 scoring matrix with a gap penalty of 12, a gap extend penalty of 4, and a frameshift gap penalty of 5.

Conservative substitutions typically include substitutions within the following groups: glycine, alanine; valine, isoleucine, leucine; aspartic acid, glutamic acid, asparagine, glutamine; serine, threonine; lysine, arginine; and phenylalanine, tyrosine.

Also provided herein are isolated nucleic acids encoding the base editor fusion proteins, vectors comprising the isolated nucleic acids, optionally operably linked to one or more regulatory domains for expressing the variant proteins, and host cells, e.g., mammalian host cells, comprising the nucleic acids, and optionally expressing the variant proteins. In some embodiments, the host cells are stem cells, e.g., hematopoietic stem cells.

In some embodiments, the fusion proteins include a linker between the DNA binding domain (e.g., ZFN, TALE, or nCas9) and the BE domains. Linkers that can be used in these fusion proteins (or between fusion proteins in a concatenated structure) can include any sequence that does not interfere with the function of the fusion proteins. In preferred embodiments, the linkers are short, e.g., 2-20 amino acids, and are typically flexible (i.e., comprising amino acids with a high degree of freedom such as glycine, alanine, and serine). In some embodiments, the linker comprises one or more units consisting of GGGS (SEQ ID NO:135) or GGGGS (SEQ ID NO:136), e.g., two, three, four, or more repeats of the GGGS (SEQ ID NO:137) or GGGGS (SEQ ID NO:138) unit. Other linker sequences can also be used.

In some embodiments, the CGBE fusion protein includes a cell-penetrating peptide sequence that facilitates delivery to the intracellular space, e.g., HIV-derived TAT peptide, penetratins, transportans, or hCT derived cell-penetrating peptides, see, e.g., Caron et al., (2001) Mol Ther. 3(3):310-8; Langel, Cell-Penetrating Peptides: Processes and Applications (CRC Press, Boca Raton Fla. 2002); E1-Andaloussi et al., (2005) Curr Pharm Des. 11(28):3597-611; and Deshayes et al., (2005) Cell Mol Life Sci. 62(16):1839-49.

Cell penetrating peptides (CPPs) are short peptides that facilitate the movement of a wide range of biomolecules across the cell membrane into the cytoplasm or other organelles, e.g. the mitochondria and the nucleus. Examples of molecules that can be delivered by CPPs include therapeutic drugs, plasmid DNA, oligonucleotides, siRNA, peptide-nucleic acid (PNA), proteins, peptides, nanoparticles, and liposomes. CPPs are generally 30 amino acids or less, are derived from naturally or non-naturally occurring protein or chimeric sequences, and contain either a high relative abundance of positively charged amino acids, e.g. lysine or arginine, or an alternating pattern of polar and non-polar amino acids. CPPs that are commonly used in the art include Tat (Frankel et al., (1988) Cell. 55:1189-1193, Vives et al., (1997) J. Biol. Chem. 272:16010-16017), penetratin (Derossi et al., (1994) J. Biol. Chem. 269:10444-10450), polyarginine peptide sequences (Wender et al., (2000) Proc. Natl. Acad. Sci. USA 97:13003-13008, Futaki et al., (2001) J. Biol. Chem. 276:5836-5840), and transportan (Pooga et al., (1998) Nat. Biotechnol. 16:857-861).

CPPs can be linked with their cargo through covalent or non-covalent strategies. Methods for covalently joining a CPP and its cargo are known in the art, e.g. chemical cross-linking (Stetsenko et al., (2000) J. Org. Chem. 65:4900-4909, Gait et al. (2003) Cell. Mol. Life. Sci. 60:844-853) or cloning a fusion protein (Nagahara et al., (1998) Nat. Med. 4:1449-1453). Non-covalent coupling between the cargo and short amphipathic CPPs comprising polar and non-polar domains is established through electrostatic and hydrophobic interactions.

CPPs have been utilized in the art to deliver potentially therapeutic biomolecules into cells. Examples include cyclosporine linked to polyarginine for immunosuppression (Rothbard et al., (2000) Nature Medicine 6(11):1253-1257), siRNA against cyclin B1 linked to a CPP called MPG for inhibiting tumorigenesis (Crombez et al., (2007) Biochem Soc. Trans. 35:44-46), tumor suppressor p53 peptides linked to CPPs to reduce cancer cell growth (Takenobu et al., (2002) Mol. Cancer Ther. 1(12):1043-1049, Snyder et al., (2004) PLoS Biol. 2:E36), and dominant negative forms of Ras or phosphoinositol 3 kinase (PI3K) fused to Tat to treat asthma (Myou et al., (2003) J. Immunol. 171:4399-4405).

CPPs have been utilized in the art to transport contrast agents into cells for imaging and biosensing applications. For example, green fluorescent protein (GFP) attached to Tat has been used to label cancer cells (Shokolenko et al., (2005) DNA Repair 4(4):511-518). Tat conjugated to quantum dots have been used to successfully cross the blood-brain barrier for visualization of the rat brain (Santra et al., (2005) Chem. Commun. 3144-3146). CPPs have also been combined with magnetic resonance imaging techniques for cell imaging (Liu et al., (2006) Biochem. and Biophys. Res. Comm. 347(1):133-140). See also Ramsey and Flynn, Pharmacol Ther. 2015 Jul. 22. pii: S0163-7258(15)00141-2.

Alternatively or in addition, the CGBE fusion proteins can include a nuclear localization sequence, e.g., SV40 large T antigen NLS (PKKKRRV (SEQ ID NO:348)) and nucleoplasmin NLS (KRPAATKKAGQAKKKK (SEQ ID NO:349)). Other NLSs are known in the art; see, e.g., Cokol et al., EMBO Rep. 2000 Nov. 15; 1(5): 411-415; Freitas and Cunha, Curr Genomics. 2009 December; 10(8): 550-557.

In some embodiments, the CGBE fusion proteins include a moiety that has a high affinity for a ligand, for example GST, FLAG or hexahistidine sequences. Such affinity tags can facilitate the purification of recombinant CGBE fusion proteins.

The CGBE fusion proteins described herein can be used for altering the genome of a cell. The methods generally include expressing or contacting the CGBE fusion proteins in the cells; in versions using one or two Cas9s, the methods include using a guide RNA having a region complementary to a selected portion of the genome of the cell. Methods for selectively altering the genome of a cell are known in the art, see, e.g., U.S. Pat. No. 8,993,233; US 20140186958; U.S. Pat. No. 9,023,649; WO/2014/099744; WO 2014/089290; WO2014/144592; WO144288; WO2014/204578; WO2014/152432; WO2115/099850; U.S. Pat. No. 8,697,359; US20160024529; US20160024524; US20160024523; US20160024510; US20160017366; US20160017301; US20150376652; US20150356239; US20150315576; US20150291965; US20150252358; US20150247150; US20150232883; US20150232882; US20150203872; US20150191744; US20150184139; US20150176064; US20150167000; US20150166969; US20150159175; US20150159174; US20150093473; US20150079681; US20150067922; US20150056629; US20150044772; US20150024500; US20150024499; US20150020223; US20140356867; US20140295557; US20140273235; US20140273226; US20140273037; US20140189896; US20140113376; US20140093941; US20130330778; US20130288251; US20120088676; US20110300538; US20110236530; US20110217739; US20110002889; US20100076057; US20110189776; US20110223638; US20130130248; US20150050699; US20150071899; US20150050699; US20150045546; US20150031134; US20150024500; US20140377868; US20140357530; US20140349400; US20140335620; US20140335063; US20140315985; US20140310830; US20140310828; US20140309487; US20140304853; US20140298547; US20140295556; US20140294773; US20140287938; US20140273234; US20140273232; US20140273231; US20140273230; US20140271987; US20140256046; US20140248702; US20140242702; US20140242700; US20140242699; US20140242664; US20140234972; US20140227787; US20140212869; US20140201857; US20140199767; US20140189896; US20140186958; US20140186919; US20140186843; US20140179770; US20140179006; US20140170753; WO/2008/108989; WO/2010/054108; WO/2012/164565; WO/2013/098244; WO/2013/176772; US 20150071899; Makarova et al., “Evolution and classification of the CRISPR-Cas systems” 9(6) Nature Reviews Microbiology 467-477 (1-23) (June 2011); Wiedenheft et al., “RNA-guided genetic silencing systems in bacteria and archaea” 482 Nature 331-338 (Feb. 16, 2012); Gasiunas et al., “Cas9-crRNA ribonucleoprotein complex mediates specific DNA cleavage for adaptive immunity in bacteria” 109(39) Proceedings of the National Academy of Sciences USA E2579-E2586 (Sep. 4, 2012); Jinek et al., “A Programmable Dual-RNA-Guided DNA Endonuclease in Adaptive Bacterial Immunity” 337 Science 816-821 (Aug. 17, 2012); Carroll, “A CRISPR Approach to Gene Targeting” 20(9) Molecular Therapy 1658-1660 (September 2012); U.S. Appl. No. 61/652,086, filed May 25, 2012; Al-Attar et al., Clustered Regularly Interspaced Short Palindromic Repeats (CRISPRs): The Hallmark of an Ingenious Antiviral Defense Mechanism in Prokaryotes, Biol Chem. (2011) vol. 392, Issue 4, pp. 277-289; Hale et al., Essential Features and Rational Design of CRISPR RNAs That Function With the Cas RAMP Module Complex to Cleave RNAs, Molecular Cell, (2012) vol. 45, Issue 3, 292-302.

For methods in which the CGBE fusion proteins are delivered to cells, the proteins can be produced using any method known in the art, e.g., by in vitro translation, or expression in a suitable host cell from nucleic acid encoding the CGBE fusion protein; a number of methods are known in the art for producing proteins. For example, the proteins can be produced in and purified from yeast, E. coli, insect cell lines, plants, transgenic animals, or cultured mammalian cells; see, e.g., Palomares et al., “Production of Recombinant Proteins: Challenges and Solutions,” Methods Mol Biol. 2004; 267:15-52. In addition, the CGBE fusion proteins can be linked to a moiety that facilitates transfer into a cell, e.g., a lipid nanoparticle, optionally with a linker that is cleaved once the protein is inside the cell. See, e.g., LaFountaine et al., Int J Pharm. 2015 Aug. 13; 494(1):180-194.

Expression Systems

To use the CGBE fusion proteins described herein, it may be desirable to express them from a nucleic acid that encodes them. This can be performed in a variety of ways. For example, the nucleic acid encoding the CGBE fusion can be cloned into an intermediate vector for transformation into prokaryotic or eukaryotic cells for replication and/or expression. Intermediate vectors are typically prokaryote vectors, e.g., plasmids, or shuttle vectors, or insect vectors, for storage or manipulation of the nucleic acid encoding the CGBE fusion for production of the CGBE fusion protein. The nucleic acid encoding the CGBE fusion protein can also be cloned into an expression vector, for administration to a plant cell, animal cell, preferably a mammalian cell or a human cell, fungal cell, bacterial cell, or protozoan cell.

To obtain expression, a sequence encoding a CGBE fusion protein is typically subcloned into an expression vector that contains a promoter to direct transcription. Suitable bacterial and eukaryotic promoters are well known in the art and described, e.g., in Sambrook et al., Molecular Cloning, A Laboratory Manual (3d ed. 2001); Kriegler, Gene Transfer and Expression: A Laboratory Manual (1990); and Current Protocols in Molecular Biology (Ausubel et al., eds., 2010). Bacterial expression systems for expressing the engineered protein are available in, e.g., E. coli, Bacillus sp., and Salmonella (Palva et al., 1983, Gene 22:229-235). Kits for such expression systems are commercially available. Eukaryotic expression systems for mammalian cells, yeast, and insect cells are well known in the art and are also commercially available.

The promoter used to direct expression of a nucleic acid depends on the particular application. For example, a strong constitutive promoter is typically used for expression and purification of fusion proteins. In contrast, when the CGBE fusion protein is to be administered in vivo for gene regulation, either a constitutive or an inducible promoter can be used, depending on the particular use of the CGBE fusion protein. In addition, a preferred promoter for administration of the CGBE fusion protein can be a weak promoter, such as HSV TK or a promoter having similar activity. The promoter can also include elements that are responsive to transactivation, e.g., hypoxia response elements, Gal4 response elements, lac repressor response element, and small molecule control systems such as tetracycline-regulated systems and the RU-486 system (see, e.g., Gossen & Bujard, 1992, Proc. Natl. Acad. Sci. USA, 89:5547; Oligino et al., 1998, Gene Ther., 5:491-496; Wang et al., 1997, Gene Ther., 4:432-441; Neering et al., 1996, Blood, 88:1147-55; and Rendahl et al., 1998, Nat. Biotechnol., 16:757-761).

In addition to the promoter, the expression vector typically contains a transcription unit or expression cassette that contains all the additional elements required for the expression of the nucleic acid in host cells, either prokaryotic or eukaryotic. A typical expression cassette thus contains a promoter operably linked, e.g., to the nucleic acid sequence encoding the CGBE fusion protein, and any signals required, e.g., for efficient polyadenylation of the transcript, transcriptional termination, ribosome binding sites, or translation termination. Additional elements of the cassette may include, e.g., enhancers, and heterologous spliced intronic signals.

The particular expression vector used to transport the genetic information into the cell is selected with regard to the intended use of the CGBE fusion protein, e.g., expression in plants, animals, bacteria, fungus, protozoa, etc. Standard bacterial expression vectors include plasmids such as pBR322 based plasmids, pSKF, pET23D, and commercially available tag-fusion expression systems such as GST and LacZ.

Expression vectors containing regulatory elements from eukaryotic viruses are often used in eukaryotic expression vectors, e.g., SV40 vectors, papilloma virus vectors, and vectors derived from Epstein-Barr virus. Other exemplary eukaryotic vectors include pMSG, pAV009/A+, pMTO10/A+, pMAMneo-5, baculovirus pDSVE, and any other vector allowing expression of proteins under the direction of the SV40 early promoter, SV40 late promoter, metallothionein promoter, murine mammary tumor virus promoter, Rous sarcoma virus promoter, polyhedrin promoter, or other promoters shown effective for expression in eukaryotic cells.

The vectors for expressing the CGBE fusion protein can include RNA Pol III promoters to drive expression of the guide RNAs, e.g., the H1, U6 or 7SK promoters. These human promoters allow for expression of CGBE fusion protein in mammalian cells following plasmid transfection.

Some expression systems have markers for selection of stably transfected cell lines such as thymidine kinase, hygromycin B phosphotransferase, and dihydrofolate reductase. High yield expression systems are also suitable, such as using a baculovirus vector in insect cells, with the gRNA encoding sequence under the direction of the polyhedrin promoter or other strong baculovirus promoters.

The elements that are typically included in expression vectors also include a replicon that functions in E. coli, a gene encoding antibiotic resistance to permit selection of bacteria that harbor recombinant plasmids, and unique restriction sites in nonessential regions of the plasmid to allow insertion of recombinant sequences.

Standard transfection methods are used to produce bacterial, mammalian, yeast or insect cell lines that express large quantities of protein, which are then purified using standard techniques (see, e.g., Colley et al., 1989, J. Biol. Chem., 264:17619-22; Guide to Protein Purification, in Methods in Enzymology, vol. 182 (Deutscher, ed., 1990)). Transformation of eukaryotic and prokaryotic cells are performed according to standard techniques (see, e.g., Morrison, 1977, J. Bacteriol. 132:349-351; Clark-Curtiss & Curtiss, Methods in Enzymology 101:347-362 (Wu et al., eds, 1983).

Any of the known procedures for introducing foreign nucleotide sequences into host cells may be used. These include the use of calcium phosphate transfection, polybrene, protoplast fusion, electroporation, nucleofection, liposomes, microinjection, naked DNA, plasmid vectors, viral vectors, both episomal and integrative, and any of the other well-known methods for introducing cloned genomic DNA, cDNA, synthetic DNA or other foreign genetic material into a host cell (see, e.g., Sambrook et al., supra). It is only necessary that the particular genetic engineering procedure used be capable of successfully introducing at least one gene into the host cell capable of expressing the CGBE fusion protein.

In methods wherein the fusion proteins include a Cas9 domain, the methods also include delivering at least one gRNA that interacts with the Cas9, or a nucleic acid that encodes a gRNA.

Alternatively, the methods can include delivering the CGBE fusion protein and guide RNA together, e.g., as a complex. For example, the CGBE fusion protein and gRNA can be can be overexpressed in a host cell and purified, then complexed with the guide RNA (e.g., in a test tube) to form a ribonucleoprotein (RNP), and delivered to cells. In some embodiments, the CGBE fusion protein can be expressed in and purified from bacteria through the use of bacterial expression plasmids. For example, His-tagged CGBE fusion protein can be expressed in bacterial cells and then purified using nickel affinity chromatography. The use of RNPs circumvents the necessity of delivering plasmid DNAs encoding the nuclease or the guide, or encoding the nuclease as an mRNA. RNP delivery may also improve specificity, presumably because the half-life of the RNP is shorter and there's no persistent expression of the nuclease and guide (as you′d get from a plasmid). The RNPs can be delivered to the cells in vivo or in vitro, e.g., using lipid-mediated transfection or electroporation. See, e.g., Liang et al. “Rapid and highly efficient mammalian cell engineering via Cas9 protein transfection.” Journal of biotechnology 208 (2015): 44-53; Zuris, John A., et al. “Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo.” Nature biotechnology 33.1 (2015): 73-80; Kim et al. “Highly efficient RNA-guided genome editing in human cells via delivery of purified Cas9 ribonucleoproteins.” Genome research 24.6 (2014): 1012-1019.

The present invention also includes the vectors and cells comprising the vectors, as well as kits comprising the proteins and nucleic acids described herein, e.g., for use in a method described herein.

Methods of Use

The base editors described herein can be used to generate transversion mutations—i.e., C-to-G mutations—in a nucleic acid sequence, e.g., in a cell, e.g., a cell in an animal (e.g., a mammal such as a human or veterinary subject), or a synthetic nucleic acid substrate. The methods include contacting the nucleic acid with a base editor as described herein. Where the base editor includes a CRISPR Cas9 or Cas12a protein, the methods further include the use of one or more guide RNAs that direct binding of the base editor to a sequence to be deaminated.

For example, the base editors described herein can be used for in vitro, in vivo or in situ directed evolution, e.g., to engineer polypeptides or proteins based on a synthetic selection framework, e.g. antibiotic resistance in E. coli or resistance to anti-cancer therapeutics being assayed in mammalian cells (e.g. CRISPR-X Hess et al, PMID: 27798611 or BE-plus systems Jiang et al, PMID: 29875396).

Tables

TABLE A Exemplary APOBEC1 proteins. This table lists (in alphabetical order) important APOBEC1 homologues. Uniprot Seq APOBEC1 accession ID orthologue Species number Version number NO: African elephant Loxodonta G3U0R4 version 30 of the entry 1 africana and version 1 of the sequence African lungfish Protopterus A0A0M3N0G8 version 4 of the entry 2 annectens and version 1 of the sequence American alligator Alligator A0A151P6M4 version 9 of the entry 3 mississippiensis and version 1 of the sequence American Anolis F1CGT0 version 16 of the entry 4 chameleon carolinensis and version 1 of the sequence American crow Corvus A0A091EQ78 version 8 of the entry 5 brachyrhynchos and version 1 of the sequence Anna's Calypte anna A0A091IIG0 version 9 of the entry 6 hummingbird and version 1 of the sequence Atlantic bottle- Tursiops A0A2U4ALA1 version 2 of the entry 7 nosed dolphin truncatus and version 1 of the sequence Barn owl Tyto alba A0A093FY71 version 6 of the entry 8 and version 1 of the sequence Black flying fox Pteropus alecto L5KGJ8 version 13 of the entry 9 and version 1 of the sequence Black snub-nosed Rhinopithecus A0A2K6KS69 version 5 of the entry 10 monkey bieti and version 1 of the sequence Beluga whale Delphlnapterus A0A2Y9NGP5 version 1 of the entry 11 leucas and version 1 of the sequence Bengalese finch Lonchura striata A0A218ULD2 version 3 of the entry 12 domestica and version 1 of the sequence Blue-fronted Amazona A0A0Q3WRD0 version 5 of the entry 13 Amazon parrot aestiva and version 1 of the sequence Bolivian squirrel Saimiri A0A2K6U925 version 5 of the entry 14 monkey boliviensis and version 1 of the boliviensis sequence Bonobo Pan paniscus A0A2R9A0R0 version 2 of the entry 15 and version 1 of the sequence Bornean Pongo Q694B3 version 60 of the entry 16 orangutan pygmaeus and version 2 of the sequence Bovine Bos taurus E1BP99 version 40 of the entry 17 and version 1 of the sequence Brandt's bat Myotis brandtii S7PYX0 version 9 of the entry 18 and version 1 of the sequence Cat Felis catus M3WB96 version 31 of the entry 19 and version 2 of the sequence Cebus capucinus Cebus capucinus A0A2K5PZC0 version 5 of the entry 20 imitator imitator and version 1 of the sequence Chimpanzee Pan troglodytes H2Q5C6 version 32 of the entry 21 and version 1 of the sequence Chinese alligator Alligator A0A1U7S7K7 version 5 of the entry 22 sinensis and version 1 of the sequence Chinese hamster Cricetulus G3I1S7 version 15 of the entry 23 griseus and version 1 of the sequence Chuck-will's- Antrostomus A0A094MFH1 version 10 of the entry 24 widow carolinensis and version 1 of the sequence Coquerel's sifaka Propithecus A0A2K6EVT9 version 5 of the entry 25 coquereli and version 1 of the sequence Crab-eating Macaca G8F4P7 version 11 of the entry 26 macaque fascicularis and version 1 of the sequence Crested ibis Nipponia A0A091V7F8 version 9 of the entry 27 nippon and version 1 of the sequence Dalmatian pelican Pelecanus A0A091SSF0 version 8 of the entry 28 crispus and version 1 of the sequence Damaraland mole Fukomys A0A091CVE5 version 9 of the entry 29 rat damarensis and version 1 of the sequence David's myotis Myotis davidii L5LUG3 version 11 of the entry 30 and version 1 of the sequence Dog Canis lupus F1PUJ5 version 41 of the entry 31 familiaris and version 2 of the sequence Downy Dryobates A0A093GVH6 version 9 of the entry 32 woodpecker pubescens and version 1 of the sequence Drill Mandrillus A0A2K5Z8Y4 version 4 of the entry 33 leucophaeus and version 1 of the sequence East African grey Balearica A0A087VMP5 version 8 of the entry 34 crowned-crane regulorum and version 1 of the gibbericeps sequence Emperor penguin Aptenodytes A0A087QNJ5 version 8 of the entry 35 forsteri and version 1 of the sequence Enhydra lutris Enhydra lutris A0A2Y9IYV0 version 1 of the entry 36 kenyoni kenyoni and version 1 of the sequence European Mustela B2NIW5 version 34 of the entry 37 domestic ferret putorius furo and version 1 of the sequence Florida manatee Trichechus A0A2Y9E587 version 1 of the entry 38 manatus and version 1 of the latirostris sequence Giant panda Ailuropoda G1LKL4 version 27 of the entry 39 melanoleuca and version 1 of the sequence Golden-collared Manacus A0A093PWR2 version 8 of the entry 40 manakin vitellinus and version 1 of the sequence Golden hamster Mesocricetus Q9EQP0 version 73 of the entry 41 auratus and version 1 of the sequence Golden snub- Rhinopithecus A0A2K6PRF3 version 4 of the entry 42 nosed monkey roxellana and version 1 of the sequence Green monkey Chlorocebus A0A0D9RBS4 version 11 of the entry 43 sabaeus and version 1 of the sequence Guinea pig Cavia porcellus A0A286XNR2 version 5 of the entry 44 and version 1 of the sequence Hawaiian monk Neomonachus A0A2Y9HAT6 version 1 of the entry 45 seal schauinslandi and version 1 of the sequence Hoatzin Opisthocomus A0A091XJL0 version 8 of the entry 46 hoazin and version 1 of the sequence Horse Equus ferus F6WR88 version 28 of the entry 47 caballus and version 1 of the sequence Human Homo sapiens P41238 version 166 of the entry 48 and version 3 of the sequence Kea Nestor notabilis A0A091RU17 version 8 of the entry 49 and version 1 of the sequence Little egret Egretta garzetta A0A091IWL9 version 10 of the entry 50 and version 1 of the sequence Ma's night Aotus A0A2K5DG70 version 6 of the entry 51 monkey nancymaae and version 1 of the sequence Mouse Mus musculus P51908 version 150 of the entry 52 and version 1 of the sequence Naked mole rat Heterocephalus G5BPM8 version 16 of the entry 53 glaber and version 1 of the sequence Northern carmine Merops nubicus A0A091QEK6 version 8 of the entry 54 bee-eater and version 1 of the sequence Northern fulmar Fulmarus A0A093LP85 version 9 of the entry 55 glacialis and version 1 of the sequence Northern white- Nomascus G1QZV0 version 31 of the entry 56 cheeked gibbon leucogenys and version 1 of the sequence Olive baboon Papio anubis A0A096MWB4 version 19 of the entry 57 and version 2 of the sequence Gray short-tailed Monodelphis Q9TUI7 version 101 of the entry 58 Opossum domestica and version 1 of the sequence Ord's kangaroo Dipodomys ordii A0A1S3FTE2 version 3 of the entry 59 rat and version 1 of the sequence Pacific walrus Odobenus A0A2U3WPA5 version 2 of the entry 60 rosmarus and version 1 of the divergens sequence Patagioenas Patagioenas A0A1V4JAP2 version 3 of the entry 61 fasciata monilis fasciata monilis and version 1 of the sequence Peters' Angolan Colobus A0A2K5JKV4 version 4 of the entry 62 colobus angolensis and version 1 of the palliatus sequence Philippine tarsier Tarsius syrichta A0A1U7U8J6 version 3 of the entry 63 and version 1 of the sequence Pig Sus scrofa F1SLW4 version 37 of the entry 64 and version 2 of the sequence Pig-tailed Macaca A0A2K6BGI5 version 4 of the entry 65 macaque nemestrina and version 1 of the sequence Rabbit Oryctolagus P47855 version 96 of the entry 66 cuniculus and version 1 of the sequence Rat Rattus P38483 version 137 of the entry 67 norvegicus and version 1 of the sequence Red-legged Cariama A0A091M4D7 version 10 of the entry 68 seriema cristata and version 1 of the sequence Red throated Gavia stellata A0A093F3R4 version 8 of the entry 69 diver and version 1 of the sequence Rhesus macaque Macaca mulatta G7N5W0 version 19 of the entry 70 and version 1 of the sequence Rifleman Acanthisitta A0A091MEP8 version 8 of the entry 71 (Acanthisitta chloris and version 1 of the chloris) sequence Rock dove Columba livia A0A2I0LXZ8 version 3 of the entry 72 and version 1 of the sequence Sheep Ovis aries W5NVH9 version 19 of the entry 73 and version 1 of the sequence Small-eared Otolemur H0XVG8 version 27 of the entry 74 galago gamettii and version 1 of the (Garnett's greater sequence bushbaby) Smooth Stylophora A0A2B4RXQ3 version 4 of the entry 75 cauliflower coral pistillata and version 1 of the sequence Sooty mangabey Cercocebus A0A2K5L2J6 version 5 of the entry 76 atys and version 1 of the sequence Sperm whale Physeter A0A2Y9T649 version 1 of the entry 77 macrocephalus and version 1 of the sequence Sumatran Pongo abelii H2NGD0 version 24 of the entry 78 orangutan and version 1 of the sequence. Sunbittern Eurypyga helias A0A093JI54 version 8 of the entry 79 and version 1 of the sequence Tasmanian devil Sarcophilus G3W4I1 version 32 of the entry 80 harrisii and version 1 of the sequence Weddell seal Leptonychotes A0A2U3Y3M5 version 2 of the entry 81 weddellii and version 1 of the sequence Western Erinaceus A0A1S3AN78 version 3 of the entry 82 European europaeus and version 1 of the hedgehog sequence White-tailed sea- Haliaeetus A0A091PSV3 version 8 of the entry 83 eagle albicilla and version 1 of the sequence White tufted ear Callithrix F7F6M6 version 31 of the entry 84 marmoset jacchus and version 2 of the sequence Wild yak Bos mutus L8IDZ0 version 15 of the entry 85 and version 1 of the sequence Yellow-throated Pterocles A0A093CIQ8 version 5 of the entry 86 sandgrouse gutturalis and version 1 of the sequence

TABLE B Exemplary APOBEC/AID family proteins. The following table lists (in alphabetical order) exemplary APOBEC family homologues. APOBEC/AID Uniprot family accession Seq. homologue number Version number ID Rat APOBEC1 P38483 version 137 of the entry 67 and version 1 of the sequence Human AID Q9GZX7 version 155 of the entry 87 (AICDA) and version 1 of the sequence Human P41238 version 166 of the entry 48 APOBEC1 and version 3 of the sequence Human Q9Y235 version 132 of the entry 88 APOBEC2 and version 1 of the sequence Human P31941 version 160 of the entry 89 APOBEC3A and version 3 of the sequence Human Q9UH17 version 150 of the entry 90 APOBEC3B and version 1 of the sequence Human Q9NRW3 version 147 of the entry 91 APOBEC3C and version 2 of the sequence Human Q96AK3 version 127 of the entry 92 APOBEC3D and version 1 of the sequence Human Q8IUX4 version 143 of the entry 93 APOBEC3F and version 3 of the sequence Human Q9HC16 version 168 of the entry 94 APOBEC3G and version 1 of the sequence Human Q6NTF7 version 115 of the entry 95 APOBEC3H and version 4 of the sequence Petromyzon NCBI Genbank: Version 1 of the entry, 96 marinus cytosine ABO15149.1 accession EF094822.1 deaminase Uniprot: A5H718 (pmCDA1) Petromyzon Same sequence — 97 marinus cytosine as ID 96, but deaminase with R187W (pmCDA1) mutation

TABLE C Exemplary TadA proteins. Some or all residues listed in Table A as well as combinations thereof might also be introduced in any of these TadA orthologues or tRNA adenosine deaminase homologues (see FIG. 5 for alignments of these TadA proteins). tRNA-specific Uniprot adenosine accession Sequence Seq. deaminase number version # ID E. coli TadA P68398 2 98 S. aureus TadA Q99W51 1 99 S. pyogenes TadA Q5XE14 2 100 S. typhi TadA Q8XGY4 2 101 A. aeolicus TadA O67050 1 102 S. pombe TAD2 O94642 2 103 S. cerevisiae TAD1 P53065 1 104 S. cerevisiae TAD2 P47058 1 105 A. thaliana TAD2 Q6IDB6 1 106 X. laevis ADAT2 Q4V7V8 1 107 X. tropicalis ADAT2 Q0P4H0 1 108 D. rerio ADAT2 Q5RIV4 2 109 B. taurus ADAT2 Q5E9J7 1 110 M. musculus ADAT2 Q6P6J0 1 111 H. sapiens ADAT2 Q7Z6V5 1 112

TABLE D Specific codons and amino acid modifications that are actionable with CGBE. Listing potential codon changes, as well as amino acid modifications that can be induced by CGBE. WT = wild type; AA = amino acid; = same AA also included in potential outcome. wt wt codon mutations, AA mutations, codon mutations, AA mutations, codon AA C-to-G C-to-G G-to-C G-to-C AAA K AAA = AAA = AAC N AAG N > K AAC = AAG K AAG = AAC K > N AAT N AAT = AAT = ACA T AGA T > R ACA = ACC T AGG, AGC, ACG T > R, T > S, = ACC = ACG T AGG T > R ACC = ACT T AGT T > S ACT = AGA R AGA = ACA R > T AGC S AGG S > R ACC S > T AGG R AGG = ACC, ACG, AGC R > T, R > S AGT S AGT = ACT S > T ATA I ATA = ATA = ATC I ATG I > M ATC = ATG M ATG = ATC M > I ATT I ATT = ATT = CAA Q GAA Q > E CAA = CAC H GAG, GAC, CAG H > E, H > D, H > Q CAC = CAG Q GAG Q > E CAC Q > H CAT H GAT H > D CAT = CCA P GGA, GCA, CGA P > G, P > A, P > R CCA = CCC P GGG, GCC, CGC, P > G, P > A, CCC = CCG, GGC, CGG, P > R, = GCG CCG P GGG, GCG, CGG P > G, P > A, P > R CCC = CCT P GGT, GCT, CGT P > G, P > A, P > R CCT = CGA R GGA R > G CCA R > P CGC R GGG, GGC, CGG R > G, = CCC R > P CGG R GGG R > G CCC, CCG, CGC R > P, = CGT R GGT R > G CCT R > P CTA L GTA L > V CTA = CTC L GTG, GTC, CTG L > V, = CTC = CTG L GTG L > V CTC = CTT L GTT L > V CTT = GAA E GAA = CAA E > Q GAC D GAG D > E CAC D > H GAG E GAG = CAC, CAG, GAC E > H, E > Q, E > D GAT D GAT = CAT D > H GCA A GGA A > G CCA A > P GCC A GGG, GGC, GCG A > G, = CCC A > P GCG A GGG A > G CCC, CCG, GCC A > P, = GCT A GGT A > G CCT A > P GGA G GGA = CCA, CGA, GCA G > P, G > R, G > A GGC G GGG = CCC, CGC, GCC G > P, G > R, G > A GGG G GGG = CCC, CGG, GCG, G > P, G > R, GGC, CCG, GCC, G > A, = CGC GGT G GGT = CCT, CGT, GCT G > P, G > R, G > A GTA V GTA = CTA V > L GTC V GTG = CTC V > L GTG V GTG = CTC, CTG, GTC V > L, = GTT V GTT = CTT V > L TAA * TAA = TAA = TAC Y TAG Y > * TAC = TAG * TAG = TAC * > Y TAT Y TAT = TAT = TCA S TGA S > * TCA = TCC S TGG, TGC, TCG S > W, S > C, = TCC = TCG S TGG S > W TCC = TCT S TGT S > C TCT = TGA * TGA = TCA * > S TGC C TGG C > W TCC C > S TGG W TGG = TCC, TCG, TGC W > S, W > C TGT C TGT = TCT C > S TTA L TTA = TTA = TTC F TTG F > L TTC = TTG L TTG = TTC L > F TTT F TTT = TTT =

TABLE E1 Specific targetable mutations from the ClinVar database that can be corrected with CGBE using Cas9 proteins with NGG-PAM recognition. snpId name geneId phenotypeList 121908088 C > G 7173|TPO Deficiency of iodide peroxidase, not provided 143367518 C > G 1161|ERCC8 Cockayne syndrome type A, not provided 74953290 C > G 324|APC Hereditary cancer-predisposing syndrome, not provided, not specified 201732356 C > G 5428|POLG not provided 587783598 C > G 1785|DNM2 Myopathy, centronuclear, not provided 879254375 C > G 3949|LDLR Familial hypercholesterolemia 752596535 C > G 3949|LDLR Familial hypercholesterolemia 121908725 C > G 100|ADA Severe combined immunodeficiency due to ADA deficiency, not provided 587777526 C > G 23394|ADNP Helsmoortel-van der aa syndrome, Inborn genetic diseases, not provided 398123527 G > C 2629|GBA Gaucher disease, Gaucher's disease, type 1 794728589 G > C 4000|LMNA Primary dilated cardiomyopathy, not provided 267607570 G > C 4000|LMNA Cardiovascular phenotype, Charcot-Marie-Tooth disease, type 2, Dilated cardiomyopathy 1A, not provided 1167218743 G > C 3030|HADHA Long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency, Long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency, Mitochondrial trifunctional protein deficiency 727504799 G > C 7273|TTN Cardiomyopathy, Primary dilated cardiomyopathy 767978961 G > C 729920|CRPPA Congenital muscular dystrophy-dystroglycanopathy with brain and eye anomalies, type A7, Muscular dystrophy-dystroglycanopathy (limb-girdle), type c, 7, not provided 1325951163 G > C 673|BRAF Global developmental delay, not provided 398123181 G > C 2592|GALT Deficiency of UDPglucose-hexose-1-phosphate uridylyltransferase, not provided 137853150 G > C 10312|TCIRG1 Osteopetrosis autosomal recessive 1 759520465 G > C 472|ATM Ataxia-telangiectasia syndrome, Hereditary cancer- predisposing syndrome, not provided 539407162 G > C 89910|UBE3B Inborn genetic diseases, Kaufman oculocerebrofacial syndrome 63750473 G > C 368|ABCC6 Pseudoxanthoma elasticum, not provided 397516354 G > C 7137|TNNI3 Hypertrophic cardiomyopathy, Primary familial hypertrophic cardiomyopathy

TABLE E2 Specific targetable mutations from the ClinVar database that can be corrected with CGBE using Cas9 proteins with NGA-PAM recognition. snpId name geneId phenotypeList 536746349 C > G 1716|DGUOK Progressive external ophthalmoplegia with mitochondrial DNA deletions, autosomal recessive 4, not provided C > G 4703|NEB Nemaline myopathy, Nemaline myopathy 2 398123350 C > G 272O|GLB1 GM1 gangliosidosis type 2, GM1 gangliosidosis type 2, Gangliosidosis GM1 type 3, Gangliosidosis GM1 type 3, Infantile GM1 gangliosidosis, Infantile GM1 gangliosidosis, Mucopolysaccharidosis, MPS-IV- B, Mucopolysaccharidosis, MPS-IV-B, not provided 121913286 C > G 5290|PIK3CA Adenocarcinoma of prostate, Adenocarcinoma of stomach, Breast adenocarcinoma, Glioblastoma, Malignant melanoma of skin, Malignant neoplasm of body of uterus, Medulloblastoma, Neoplasm of brain, Neoplasm of the breast, Neoplasm of the large intestine, Squamous cell carcinoma of the head and neck, Transitional cell carcinoma of the bladder, Uterine Carcinosarcoma, Uterine cervical neoplasms C > G 5896|RAG1 Alpha/beta T-cell lymphopenia with gamma/delta T-cell expansion, severe cytomegalovirus infection, and autoimmunity, Combined cellular and humoral immune defects with granulomas, Combined cellular and humoral immune defects with granulomas, Histiocytic medullary reticulosis, Severe immunodeficiency, autosomal recessive, T-cell negative, B-cell negative, NK cell- positive, Severe immunodeficiency, autosomal recessive, T- cell negative, B-cell negative, NK cell-positive 1057517774 C > G 4647|MYO7A Deafness, autosomal recessive 2, Usher syndrome, type 1, not provided 749491616 C > G 35|ACADS Deficiency of butyryl-CoA dehydrogenase, not provided 761649878 C > G 5428|POLG POLG-Related disorder, Progressive sclerosing poliodystrophy, not provided 104894718 C > G 6324|SCN1B Atrial fibrillation, familial, 13, Atrial fibrillation, familial, 13, Brugada syndrome 5, Brugada syndrome 5, Epileptic encephalopathy, early infantile, 52, Generalized epilepsy with febrile seizures plus, Generalized epilepsy with febrile seizures plus, type 1, Generalized epilepsy with febrile seizures plus, type 1, Seizures, not provided 876657730 G > C 7399|USH2A Retinitis pigmentosa 39, Usher syndrome, type 2A, Usher syndrome, type 2A, not provided 869320742 G > C 7273|TTN Hereditary myopathy with early respiratory failure, not provided 672601366 G > C 547|KIF1A Mental retardation, autosomal dominant 9 863224905 G > C 64324|NSD1 Beckwith-Wiedemann syndrome, Sotos syndrome 1 756013171 G > C 157680|VPS13B Cohen syndrome, not provided 120074186 G > C 3784|KCNQ1 Cardiovascular phenotype, Congenital long QT syndrome, Jervell and Lange-Nielsen syndrome 1, not provided 121908195 G > C 1200|TPP1 Ceroid lipofuscinosis neuronal 2, not provided 1057517420 G > C 6833|ABCC8 Familial hyperinsulinism, Persistent hyperinsulinemic hypoglycemia of infancy 81002840 G > C 675|BRCA2 Familial cancer of breast, Hereditary breast and ovarian cancer syndrome, not provided 730882218 G > C 4247|MGAT2 Abnormal facial shape, Abnormal glycosylation (CDG IIa), Carbohydrate-deficient glycoprotein syndrome type II, Global developmental delay 1555534596 G > C 4763|NF1 Hereditary cancer-predisposing syndrome, not provided 80358254 G > C 4864|NPC1 Niemann-Pick disease type C1, Niemann-Pick disease, type C, not provided G > C 6261|RYR1 not provided 147484110 G > C 1476|CSTB Epilepsy, progressive myoclonic 1A (Unverricht and Lundborg), Inborn genetic diseases, Progressive myoclonic epilepsy, Unverricht-Lundborg syndrome, not provided

TABLE E3 Specific targetable mutations from the ClinVar database that can be corrected with CGBE using Cas9 proteins with NG-PAM recognition. snpId name geneId phenotypeList 370124822 C > G 4595|MUTYH Hereditary cancer-predisposing syndrome, MYH-associated polyposis, not provided 121908088 C > G 7173|TPO Deficiency of iodide peroxidase, not provided 536746349 C > G 1716|DGUOK Progressive external ophthalmoplegia with mitochondrial DNA deletions, autosomal recessive 4, not provided C > G 4703|NEB Nemaline myopathy, Nemaline myopathy 2 557312035 C > G 7273|TTN Dilated cardiomyopathy 1G, Limb-girdle muscular dystrophy, type 2J, Primary dilated cardiomyopathy 587781707 C > G 580|BARD1 Breast cancer, susceptibility to, Familial cancer of breast, Hereditary cancer-predisposing syndrome, not provided 398123350 C > G 272O|GLB1 GM1 gangliosidosis type 2, GM1 gangliosidosis type 2, Gangliosidosis GM1 type 3, Gangliosidosis GM1 type 3, Infantile GM1 gangliosidosis, Infantile GM1 gangliosidosis, Mucopolysaccharidosis, MPS-IV- B, Mucopolysaccharidosis, MPS-IV-B, not provided 121913286 C > G 5290|PIK3CA Adenocarcinoma of prostate, Adenocarcinoma of stomach, Breast adenocarcinoma, Glioblastoma, Malignant melanoma of skin, Malignant neoplasm of body of uterus, Medulloblastoma, Neoplasm of brain, Neoplasm of the breast, Neoplasm of the large intestine, Squamous cell carcinoma of the head and neck, Transitional cell carcinoma of the bladder, Uterine Carcinosarcoma, Uterine cervical neoplasms 143367518 C > G 1161|ERCC8 Cockayne syndrome type A, not provided 74953290 C > G 324|APC Hereditary cancer-predisposing syndrome, not provided, not specified 730881857 C > G 4683|NBN Hereditary cancer-predisposing syndrome, Microcephaly, normal intelligence and immunodeficiency, not provided 878853697 C > G 2705|GJB1 Charcot-Marie-Tooth Neuropathy X, not provided 33941377 C > G 3043|HBB Beta thalassemia intermedia, Beta-plus-thalassemia, beta Thalassemia, not provided C > G 5896|RAG1 Alpha/beta T-cell lymphopenia with gamma/delta T-cell expansion, severe cytomegalovirus infection, and autoimmunity, Combined cellular and humoral immune defects with granulomas, Combined cellular and humoral immune defects with granulomas, Histiocytic medullary reticulosis, Severe immunodeficiency, autosomal recessive, T- cell negative, B-cell negative, NK cell-positive, Severe immunodeficiency, autosomal recessive, T-cell negative, B-cell negative, NK cell-positive 397515905 C > G 4607|MYBPC3 Cardiovascular phenotype, Familial hypertrophic cardiomyopathy 1, Hypertrophic cardiomyopathy, Primary familial hypertrophic cardiomyopathy, not provided 1057517774 C > G 4647|MYO7A Deafness, autosomal recessive 2, Usher syndrome, type 1, not provided 587779833 C > G 472|ATM Ataxia-telangiectasia syndrome, Ataxia-telangiectasia syndrome, Familial cancer of breast, Hereditary cancer- predisposing syndrome, not provided 137853043 C > G 7846|TUBA1A Tubulinopathies, not provided 1057520574 C > G 7846|TUBA1A Tubulinopathies, not provided 749491616 C > G 35|ACADS Deficiency of butyryl-CoA dehydrogenase, not provided 201732356 C > G 5428|POLG not provided 761649878 C > G 5428|POLG POLG-Related disorder, Progressive sclerosing poliodystrophy, not provided 769410130 C > G 5428|POLG Progressive sclerosing poliodystrophy, not provided 587783598 C > G 1785|DNM2 Myopathy, centronuclear, not provided 879254375 C > G 3949|LDLR Familial hypercholesterolemia 752596535 C > G 3949|LDLR Familial hypercholesterolemia 875989909 C > G 3949|LDLR Familial hypercholesterolemia, Familial hypercholesterolemias 104894718 C > G 6324|SCN1B Atrial fibrillation, familial, 13, Atrial fibrillation, familial, 13, Brugada syndrome 5, Brugada syndrome 5, Epileptic encephalopathy, early infantile, 52, Generalized epilepsy with febrile seizures plus, Generalized epilepsy with febrile seizures plus, type 1, Generalized epilepsy with febrile seizures plus, type 1, Seizures, not provided 121908725 C > G 100|ADA Severe combined immunodeficiency due to ADA deficiency, not provided 587777526 C > G 23394|ADNP Helsmoortel-van der aa syndrome, Inborn genetic diseases, not provided 869312901 G > C 6497|SKI Shprintzen-Goldberg syndrome, not provided 397516833 G > C 6390|SDHB Gastrointestinal stroma tumor, Hereditary Paraganglioma- Pheochromocytoma Syndromes, Hereditary cancer- predisposing syndrome, Paragangliomas 4, Paragangliomas 4, Pheochromocytoma, not provided 398123527 G > C 2629|GBA Gaucher disease, Gaucher's disease, type 1 794728589 G > C 4000|LMNA Primary dilated cardiomyopathy, not provided 267607570 G > C 4000|LMNA Cardiovascular phenotype, Charcot-Marie-Tooth disease, type 2, Dilated cardiomyopathy 1A, not provided 397517977 G > C 7399|USH2A Retinitis pigmentosa 39, Usher syndrome, type 2A, Usher syndrome, type 2A 876657730 G > C 7399|USH2A Retinitis pigmentosa 39, Usher syndrome, type 2A, Usher syndrome, type 2A, not provided 1167218743 G > C 3030|HADHA Long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency, Long-chain 3-hydroxyacyl-CoA dehydrogenase deficiency, Mitochondrial trifunctional protein deficiency 869320742 G > C 7273|TTN Hereditary myopathy with early respiratory failure, not provided 727504799 G > C 7273|TTN Cardiomyopathy, Primary dilated cardiomyopathy 672601366 G > C 547|KIF1A Mental retardation, autosomal dominant 9 587784141 G > C 64324|NSD1 Beckwith-Wiedemann syndrome, Sotos syndrome 1 863224905 G > C 64324|NSD1 Beckwith-Wiedemann syndrome, Sotos syndrome 1 988423880 G > C 5395|PMS2 Hereditary cancer-predisposing syndrome, Lynch syndrome 767978961 G > C 729920|CRPPA Congenital muscular dystrophy-dystroglycanopathy with brain and eye anomalies, type A7, Muscular dystrophy- dystroglycanopathy (limb-girdle), type c, 7, not provided 1325951163 G > C 673|BRAF Global developmental delay, not provided 756013171 G > C 157680|VPS13B Cohen syndrome, not provided 398123181 G > C 2592|GALT Deficiency of UDPglucose-hexose-1-phosphate uridylyltransferase, not provided 137853022 G > C 8518|ELP1 Familial dysautonomia 104894845 G > C 2717|GLA Fabry disease, not provided 104894229 G > C HRAS, LRRC56 Neoplasm of the large intestine, Neoplasm of the thyroid gland 120074186 G > C 3784|KCNQ1 Cardiovascular phenotype, Congenital long QT syndrome, Jervell and Lange-Nielsen syndrome 1, not provided 121908195 G > C 1200|TPP1 Ceroid lipofuscinosis neuronal 2, not provided 1057517420 G > C 6833|ABCC8 Familial hyperinsulinism, Persistent hyperinsulinemic hypoglycemia of infancy 748523268 G > C 582|BBS1 Bardet-Biedl syndrome 1 137853150 G > C 10312|TCIRG1 Osteopetrosis autosomal recessive 1 876659710 G > C 472|ATM Ataxia-telangiectasia syndrome, Hereditary cancer- predisposing syndrome 759520465 G > C 472|ATM Ataxia-telangiectasia syndrome, Hereditary cancer- predisposing syndrome, not provided 539407162 G > C 89910|UBE3B Inborn genetic diseases, Kaufman oculocerebrofacial syndrome 199474813 G > C 4633|MYL2 Familial hypertrophic cardiomyopathy 10, not provided 81002840 G > C 675|BRCA2 Familial cancer of breast, Hereditary breast and ovarian cancer syndrome, not provided 80358871 G > C 675|BRCA2 Breast-ovarian cancer, familial 2, Hereditary cancer- predisposing syndrome, not provided 730882218 G > C 4247|MGAT2 Abnormal facial shape, Abnormal glycosylation (CDG IIa), Carbohydrate-deficient glycoprotein syndrome type II, Global developmental delay 2229311 G > C 3712|IVD Isovaleryl-CoA dehydrogenase deficiency 778768583 G > C 825|CAPN3 Limb-girdle muscular dystrophy, type 2A, not provided 63750473 G > C 368|ABCC6 Pseudoxanthoma elasticum, not provided 912983346 G > C 6687|SPG7 Hereditary spastic paraplegia, not provided 587778720 G > C 7157|TP53 Adenocarcinoma of prostate, Adenocarcinoma of stomach, Adenoid cystic carcinoma, Adrenocortical carcinoma, Carcinoma of esophagus, Glioblastoma, Hepatocellular carcinoma, Hereditary cancer-predisposing syndrome, Lung adenocarcinoma, Malignant melanoma of skin, Malignant neoplasm of body of uterus, Nasopharyngeal Neoplasms, Neoplasm of brain, Neoplasm of the breast, Neoplasm of the large intestine, Ovarian Serous Cystadenocarcinoma, Pancreatic adenocarcinoma, Renal cell carcinoma, papillary, 1, Squamous cell carcinoma of the head and neck, Squamous cell carcinoma of the skin, Squamous cell lung carcinoma, Transitional cell carcinoma of the bladder, Uterine Carcinosarcoma, not specified 1555534596 G > C 4763|NF1 Hereditary cancer-predisposing syndrome, not provided 80358010 G > C 672|BRCA1 Breast-ovarian cancer, familial 1, Hereditary cancer- predisposing syndrome 80358254 G > C 4864|NPCI Niemann-Pick disease type C1, Niemann-Pick disease, type C, not provided 200727689 G > C 3949|LDLR Familial hypercholesterolemia 879254565 G > C 3949|LDLR Familial hypercholesterolemia 879254729 G > C 3949|LDLR Familial hypercholesterolemia 121908036 G > C 3949|LDLR Familial hypercholesterolemia 28942082 G > C 3949|LDLR Familial hypercholesterolemia 875989926 G > C 3949|LDLR Familial hypercholesterolemia G > C 6261|RYR1 not provided 398123508 G > C 593|BCKDHA Maple syrup urine disease, not provided 397516354 G > C 7137|TNNI3 Hypertrophic cardiomyopathy, Primary familial hypertrophic cardiomyopathy 147484110 G > C 1476|CSTB Epilepsy, progressive myoclonic 1A (Unverricht and Lundborg), Inborn genetic diseases, Progressive myoclonic epilepsy, Unverricht-Lundborg syndrome, not provided

TABLE F List of Exemplary Cas9 or Cas12a Orthologs UniProt or GenBank Nickase Accession Mutations/Catalytic Ortholog Number residues S. pyogenes Cas9 Q99ZW2.1 D10A, E762A, H840A, (SpCas9) N854A, N863A, D986A¹⁷ S. aureus Cas9 J7RUA5.1 D10A and N580¹⁸ (SaCas9) S. thermophilus Cas9 G3ECR1.2 D31A and N891A¹⁹ (St1Cas9) S. pasteurianus Cas9 BAK30384.1 D10, H599* (SpaCas9) C. jejuni Cas9 Q0P897.1 D8A, H559A²⁰ (CjCas9) F. novicida Cas9 A0Q5Y3.1 D11, N995²¹ (FnCas9) P. lavamentivorans A7HP89.1 D8, H601* Cas9 (PlCas9) C. lari Cas9 (ClCas9) G1UFN3.1 D7, H567* Pasteurella multocida Q9CLT2.1 Cas9 F. novicida Cpf1 A0Q7Q2.1 D917, E1006, D1255²¹ (FnCpf1) M. bovoculi Cpf1 WP 052585281.1 D986A** (MbCpf1) A. sp. BV3L6 Cpf1 U2UMQ6.1 D908, 993E, Q1226, D1263²³ (AsCpf1) L. bacterium N2006 A0A182DWE3.1 D832A²⁴ (LbCpf1) *predicted based on UniRule annotation on the UniProt database. **Unpublished but deposited at addgene by Ervin Welker: pTE4565 (Addgene plasmid # 88903)

TABLE G List of Exemplary High Fidelity and/or PAM-relaxed RGN Orthologs Published HF/PAM-RGN variants PMID Mutations* S. pyogenes Cas9 26628643 K810A/K1003A/R1060A (1.0); (SpCas9) eSpCasS K848A/K1003A/R1060A(1.1) S. pyogenes Cas9 29431739 M495V/Y515N/K526E/R661Q; (SpCas9) evoCas9 (M495V/Y515N/K526E/R661S; M495V/Y515N/K526E/R661L) S. pyogenes Cas9 26735016 N497A/R661A/Q695A/Q926A (SpCas9) HF1 S. pyogenes Cas9 30082871 R691A (SpCas9) HiFi Cas9 S. pyogenes Cas9 28931002 N692A, M694A, Q695A, H698A (SpCas9) HypaCas9 S. pyogenes Cas9 30082838 F539S, M763I, K890N (SpCas9) Sniper-Cas9 S. pyogenes Cas9 29512652 A262T, R324L, S409I, E480K, E543D, (SpCas9) xCas9 M694I, E1219V S. pyogenes Cas9 30166441 R1335V, L1111R, D1135V, G1218R, (SpCas9) SpCas9-NG E1219F, A1322R, T1337R S. pyogenes Cas9 26098369 D1135V, R1335Q, T1337R; (SpCas9) VQR/VRER D1135V/G1218R/R1335E/T1337R S. aureus Cas9 26524662 E782K/N968K/R1015H (SaCas9)-KKH enAsCas12a U.S. Ser. No. One or more of: E174R, S170R, S542R, 15/960,271 K548R, K548V, N551R, N552R, K607R, K607H, e.g., E174R/S542R/K548R, E174R/S542R/K607R, E174R/S542R/K548V/N552R, S170R/S542R/K548R, S170R/E174R, E174R/S542R, S170R/S542R, E174R/S542R/K548R/N551R, E174R/S542R/K607H, S170R/S542R/K607R, or S170R/S542R/K548V/N552R enAsCas12a-HF U.S. Ser. No. One or more of: E174R, S542R, K548R, 15/960,271 e.g., E174R/S542R/K548R, E174R/S542R/K607R, E174R/S542R/K548V/N552R, S170R/S542R/K548R, S170R/E174R, E174R/S542R, S170R/S542R, E174R/S542R/K548R/N551R, E174R/S542R/K607H, S170R/S542R/K607R, or S170R/S542R/K548V/N552R, with the addition of one or more of: N282A, T315A, N515A and K949A enLbCas12a(HF) U.S. Ser. No. One or more of T152R, T152K, D156R, 15/960,271 D156K, Q529K, G532R, G532K, G532Q, K538R, K538V, D541R, Y542R, M592A, K595R, K595H, K595S or K595Q, e.g., D156R/G532R/K538R, D156R/G532R/K595R, D156R/G532R/K538V/Y542R, T152R/G532R/K538R, T152R/D156R, D156R/G532R, T152R/G532R, D156R/G532R/K538R/D541R, D156R/G532R/K595H, T152R/G532R/K595R, T152R/G532R/K538V/Y542R, optionally with the addition of one or more of: N260A, N256A, K514A, D505A, K881A, S286A, K272A, K897A enFnCas12a(HF) U.S. Ser. No. One or more of T177A, K180R, K180K, 15/960,271 E184R, E184K, T604K, N607R, N607K, N607Q, K613R, K613V, D616R, N617R, M668A, K671R, K671H, K671S, or K671Q, e.g., E184R/N607R/K613R, E184R/N607R/K671R, E184R/N607R/K613V/N617R, K180R/N607R/K613R, K180R/E184R, E184R/N607R, K180R/N607R, E184R/N607R/K613R/D616R, E184R/N607R/K671H, K180R/N607R/K671R, K180R/N607R/K613V/N617R, optionally with the addition of one or more of: N305A, N301A, K589A, N580A, K962A, S334A, K320A, K978A chimeric Cas9 30718489 S. aureus Cas9 with PAM interaction cCas9 domain from SaCas9 orthologues, expands recognition and targetability of NNVRRN, NNVACT, NNVATG, NNVATT, NNVGCT, NNVGTG, and NNVGTT PAM sequences Streptococcus doi: https://doi.org/ Recognizes 5′-NAA-3′ PAM macacae (Smac) Cas9 10.1101/429654 NCTC 11558 Spy-mac Cas9, doi: https://doi.org/ Recognizes 5′-NAA-3′ PAM Smac-py Cas9 10.1101/429654 N. meningitidis 30581144 Recognizes N₄CC PAM Nme2Cas9 SpG Cas9 32217751 SpCas9 variant capable of targeting (SEQ-ID 158) NGN PAMs D1135L/S1136W/G1218K/E1219Q/ R1335Q/T1337R Also as SpG-HF1 in combination with N497A/R661A/Q695A/Q926A SpRY Cas9 32217751 SpCas9 variant capable of targeting (SEQ-ID 157) NRN > NYN PAMs SpRY(A61R/L1111R/D1135L/S1136W/ G1218K/E1219Q/N1317R/A1322R/ R1333P/R1335Q/T1337R); also as SpRY-HF1 in combination with N497A/R661A/Q695A/Q926A *predicted based on UniRule annotation on the UniProt database.

TABLE H Amino acid substitutions predicted to generate ABE variants with reduced RNA editing. This table lists the residue changes in either or both TadA domains of the TadA heterodimer (present in e.g., ABE7.10) predicted to cause an RRE phenotype, next to the reasoning behind the proposed changes. Residues to Change Rationale Wild type Engineered Protein Binding (WT) TadA TadA structure prediction S7 S205 x H8 H206 x E9 E207 x Y10 Y208 x W11 W209 x M12 M210 x R13 R211 x x H14 H212 x T17 T215 x K20 K218 x x R21 R219 x x W23 R221 x E25 E223 x x R26 R224 x x E27 E225 x V28 V226 x x P29 P227 x V30 V228 x G31 G229 x H36 L234 x N37 N235 x N38 N236 x N46 N244 x R47 R245 x P48 A246 x I49 I247 x G50 G248 x R51 I249 x H52 H250 x D53 D251 x P54 P252 x T55 T253 x A56 A254 x H57 H255 x x A58 A256 x E59 E257 x R64 R262 x Q65 Q263 x G67 G265 x L68 L266 x Q71 Q269 x N72 N270 x R74 R272 x I76 I274 x D77 D275 x Y81 Y279 x V82 V280 x T83 T281 x L84 F282 x E85 E283 x P86 P284 x x C87 C285 x x V88 V286 x M89 M287 x C90 C288 x x R98 R296 x G100 G298 x R101 R299 x A106 V304 x R107 R305 x D108 N306 x A109 A307 x K110 K308 x T111 T309 x D119 D317 x H122 H320 x H123 Y321 x P124 P322 x G125 G323 x M126 M324 x N127 N325 x H128 H326 x R129 R327 x V130 V328 x E131 E329 x I132 I330 x T133 T331 x E134 E332 x G135 G333 x L137 L335 x A138 A336 x x D139 D337 x E140 E338 x C141 C339 x x A142 A340 x x A143 A341 x x L144 L342 x L145 L343 x x S146 C344 x D147 Y345 x F148 F346 x x F149 F347 x x R150 R348 x x M151 M349 x R152 P350 x x R153 R351 x Q154 Q352 x E155 V353 x x I156 F354 x K157 N355 x K160 K358 x K161 K359 x

TABLE I Amino acid residues whose mutation may be expected to yield base editor RRE variants. These positions were chosen based on a APOBEC1 structural model and RNA/DNA binding predictions or based on previous description in the literature as residues whose mutation reduced the RNA editing or binding activities of isolated APOBEC1. Residue Change Reasoning E24, V25 model & RNA binding prediction R118, Y120, H121, R126 model & RNA binding prediction W224-K229 model & RNA binding prediction P168-I186 model & RNA binding prediction L173 + L180 model & RNA binding prediction R15, R16, R17, to K15-17 & A15-17 Teng et al, J Lipid Research 1999 Deletion E181-L210 Teng et al, J Lipid Research 1999 P190 + P191 Teng et al, J Lipid Research 1999 Deletion L210-K229 (C-terminal) Teng et al, J Lipid Research 1999 Deletion S2-L14 (N-terminal) Teng et al, J Lipid Research 1999 V64, F66 Teng et al, J Lipid Research 1999 L180A Teng et al, J Lipid Research 1999 C192, L193, L196, P201, L203, Teng et al, J Lipid Research 1999 L210, P219, P220 P92 MacGinnitie et al, JBC 1995

TABLE J UNG and SMUG analogues Uniprot accession number UNG orthologue Mouse P97931 SEQ ID NO: 318 Rat Q5BK44 SEQ ID NO: 319 Baker's yeast P12887 SEQ ID NO: 320 Caenorhabditis Q9U221 SEQ ID NO: 321 elegans Mouse-ear cress Q9LIH6 SEQ ID NO: 322 Zebrafish Q7ZVD1 SEQ ID NO: 323 Rabbit G1SJ42 SEQ ID NO: 324 Polar bear A0A452THE0 SEQ ID NO: 325 Black snub-nosed A0A2K6MB33 SEQ ID NO: 326 monkey Common wombat A0A4X2KC02 SEQ ID NO: 327 Mycobacterium A0A1X2AUJ0 SEQ ID NO: 328 riyadhense Indian major carp A0A498LRM7 SEQ ID NO: 329 Fission yeast O74834 SEQ ID NO: 330 Japanese pufferfish A0A3B5KG53 SEQ ID NO: 331 Thirteen-lined ground I3M8Q6 SEQ ID NO: 332 squirrel Japanese rice fish A0A3P9H4T8 SEQ ID NO: 333 Electric eel A0A4W4HK79 SEQ ID NO: 334 Western clawed frog A0A5G3K4Q6 SEQ ID NO: 335 Enterobacter cloacae A0A0F0TTY1 SEQ ID NO: 336 subsp, cloacae Clostridium oryzae A0A1V4IJH4 SEQ ID NO: 337 Lactobacillus apis A0A1C3ZIJ7 SEQ ID NO: 338 Flavobacterium sp. A0A519N079 SEQ ID NO: 339 Delftia lacustris A0A1H3TI78 SEQ ID NO: 340 Lactococcus garvieae A0A3D4RH89 SEQ ID NO: 341 Lactobacillus rodentium A0A2Z6T8A7 SEQ ID NO: 342 SMUG orthologue Human Q53HV7 SEQ ID NO: 343 Rat Q811Q1 SEQ ID NO: 344 Mouse Q6P5C5 SEQ ID NO: 345 African clawed frog Q9YGN6 SEQ ID NO: 346 Bovine Q59I47 SEQ ID NO: 347

EXAMPLES

The invention is further described in the following examples, which do not limit the scope of the invention described in the claims.

Methods Molecular Cloning

All base editor (BE) and prime editor (PE) constructs were cloned into a mammalian expression plasmid backbone under the control of a pCMV promoter (AgeI and NotI restriction digest of parental plasmid Addgene #112101). The wild-type SpCas9 construct (SQT 817; Addgene #53373) is expressed under the control of a CAG promoter. All BE and PE constructs were encoded as P2A-eGFP fusions for co-translational expression of the base/prime editors and eGFP. Gibson fragments with matching overlaps were PCR-amplified using Phusion High-fidelity polymerase (NEB). Fragments were gel-purified and assembled for 1 hour at 50° C. and transformed into chemically competent E. coli (XL1-Blue, Agilent). The UNGs used in our experiments originated either from E. coli (eUNG; UniProtKB-P12295) or Homo sapiens (hUNG; UniProtKB-P13051), were codon-optimized for expression in human cells and synthesized as gblocks (IDT). All guide RNA (gRNA) constructs were cloned into a BsmBI-digested pUC19-based entry vector (BPK1520, Addgene #65777) with a U6 promoter driving gRNA expression. We designed the pegRNAs to implement the same C-to-G changes that the CGBE constructs would install and followed previously described default design rules for designing pegRNAs and ngRNAs15. PegRNAs were cloned into the BsaI-digested pU6-pegRNA-GG-acceptor entry vector (Addgene #132777) and ngRNAs were cloned into the abovementioned BsmBI-digested entry vector BPK1520. Oligos containing the spacer, the 5′phosphorylated pegRNA scaffold, and the 3′ extension sequences were annealed to form dsDNA fragments with compatible overhangs and ligated using T4 ligase (NEB). All plasmids used for transfection experiments were prepared using Qiagen Midi or Maxi Plus kits.

Guide RNAs

All gRNAs for base editors were of the form (SEQ ID NO 145) 5′-NNNNNNNNNNNNNNNNNNNNCGTTTTAGAGCTAGAAATAGCAAGTT AAAATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCACCGAGTCGGT GCTTTTTTT-3′. 

TABLE K Shown below are the protospacer regions (NNNNNNNNNNNNNNNNNNNN in SEQ ID NO: 146) for these gRNAs (all written 5′ to 3′). target gene/site protospacer sequence SEQ ID NO: ABE site 7 GAATACTAAGCATAGACTCC 216 ABE site 8 GTAAACAAAGCATAGACTGA 217 ABE site 9 GAAGACCAAGGATAGACTGC 218 ABE site 18 ACACACACACTTAGAATCTG 219 ABE site 19 CACACACACTTAGAATCTGT 220 ABE site 20 TTAAGCTGTAGTATTATGAA 221 ABE site 21 CCTGGCCTGGGTCAATCCTT 222 EMX1 site 1 GAGTCCGAGCAGAAGAAGAA 223 EMX1 site 2 GTATTCACCTGAAAGTGTGC 224 FANCF site 1 GGAATCCCTTCTGCAGCACC 225 HEK site 2 (ABE site 1) GAACACAAAGCATAGACTGC 226 HEK site 3 GGCCCAGACTGAGCACGTGA 227 HEK site 4 GGCACTGCGGCTGGAGGTGG 228 HEK site 5 CTGGCCTGGGTCAATCCTTG 229 HEK site 6 CAAAGCAGGATGACAGGCAG 230 PDCD1 site 2 ACTTCCACATGAGCGTGGTC 231 PPP1R12C site 2 GGCACTCGGGGGCGAGAGGA 232 PPP1R12C site 3 GAGCTCACTGAACGCTGGCA 233 PPP1R12C site 4 GACCCTCAGCCGTGCTGCTC 234 PPP1R12C site 5 GCTGACTCAGAGACCCTGAG 235 PPP1R12C site 6 GGGGCTCAACATCGGAAGAG 236 PPP1R12C site 7 GCTGGCTCAGGTTCAGGAGA 237 PPP1R12C site 8 CTGCTCGGGGTGGGACTCTG 238 RNF2 site 1 GTCATCTTAGTCATTACCTG 239 VEGFA site 4 GAGGACGTGTGTGTCTGTGT 240 For C5, 7, 8 guides ABE site 23 TAAGCATAGACTCCAGGATA 241 ABE site 24 TACTCTGAGTGTACAAAAGA 242 ABE site 25 AGTAAACAAAGCATAGACTG 243 ABE site 26 TTTGTGCAAACACAGATTGC 244 ABE site 27 CGGGCATCAGAATTCCCTGG 245 EMX1 site 3 AAAGTACAAACGGCAGAAGC 246 EMX1 site 4 GTACAAACGGCAGAAGCTGG 247 FANCF site 2 GCTGCAGAAGGGATTCCATG 248 FANCF site 3 CGCCGTCTCCAAGGTGAAAG 249 FANCF site 4 AGCGATCCAGGTGCTGCAGA 250 HEK site 7 GGAACACAAAGCATAGACTG 251 HEK site 8 TGTGTTCCAGTTTCCTTTAC 252 HEK site 9 TTGTTTGCAGCTATTCAGGC 253 PPP1R12C site 9 AAGTCGAGGGAGGGATGGTA 254 PPP1R12C site 10 GACACGTGGATTGTGCTGTC 255 PPP1R12C site 11 GTCATACACTGGGCTGGCCA 256 PPP1R12C site 12 CAAAGTCCAGGACCGGCTGG 257 PPP1R12C site 13 GCATGGCTCTAGTGCTTTCC 258 PPP1R12C site 14 GGTCATACACTGGGCTGGCC 259 PPP1R12C site 15 AAGGAGACAAAGTCCAGGAC 260 PPP1R12C site 16 GATTGTGCTGTCAGGAGCTC 261 RNF2 site 2 ATGACTAAGATGACTGCCAA 262 RNF2 site 3 TGAGTTACAACGAACACCTC 263 For guides with NGT or NGAG PAM CGBE_NG site 1 ACCATCTTTTGTACACTCAG 264 CGBE_NG site 2 CACTTCTCTTCCTGCCCTCT 265 CGBE_NG site 3 (EMX1) AGCTTCTGCCGTTTGTACTT 266 CGBE_NG site 4 (RNF2) CGTCTCATATGCCCCTTGGC 267 CGBE_NG site 5 ATAGACTCCAGGATAAGGTA 268 CGBE_NG site 6 CTCAACATCGGAAGAGGGGA 269 (PPP1R12C) CGBE_VRQR site 1 TCAATCCTTGGGGCCCAGAC 270 CGBE_VRQR site 2 ATGTTCCAATCAGTACGCAG 271 (FANCF) CGBE_VRQR site 3 GATGACTGCCAAGGGGCATA 272 (RNF2) CGBE_VRQR site 4 AAGTACAAGCACTCAATGTG 273 CGBE_VRQR site 5 ACACACACTTAGAATCTGTG 274 CGBE_VRQR site 6 GCGGACAGTGGACGCGGCGG 275 (VEGFA)

TABLE L Shown below are the sequence for DNA off-target  sites (all written 5′ to 3′). SEQ ID target site sequence NO: HEK site 2 off 1 GAACACAATGCATAGATTGC 276 HEK site 2 off 2 AAACATAAAGCATAGACTGC 277 HEK site 3 off 1 CACCCAGACTGAGCACGTGC 278 HEK site 3 off 2 GACACAGACTGGGCACGTGA 279 HEK site 3 off 3 AGCTCAGACTGAGCAAGTGA 280 HEK site 3 off 4 AGACCAGACTGAGCAAGAGA 281 HEK site 3 off 5 GAGCCAGAATGAGCACGTGA 282 HEK site 4 off 1 TGCACTGCGGCCGGAGGAGG 283 HEK site 4 off 2 GGCTCTGCGGCTGGAGGGGG 284 HEK site 4 off 3 GGCACGACGGCTGGAGGTGG 285 HEK site 4 off 4 GGCATCACGGCTGGAGGTGG 286 HEK site 4 off 5 GGCGCTGCGGCGGGAGGTGG 287 EMX1 site 1 off 1 GAGTCTAAGCAGAAGAAGAA 288 EMX1 site 1 off 2 GAGGCCGAGCAGAAGAAAGA 289 EMX1 site 1 off 3 GAGTCCTAGCAGGAGAAGAA 290 EMX1 site 1 off 4 GAGTCCGGGAAGGAGAAGAA 291 EMX1 site 1 off 5 GAGCCGGAGCAGAAGAAGGA 292 FANCF site 1 off 1 GGAACCCCGTCTGCAGCACC 293 FANCF site 1 off 2 GGAGTCCCTCCTACAGCACC 294 FANCF site 1 off 3 AGAGGCCCCTCTGCAGCACC 295 FANCF site 1 off 4 ACCATCCCTCCTGCAGCACC 296 FANCF site 1 off 5 GGATTGCCATCCGCAGCACC 297 FANCF site 1 off 6 TGAATCCCATCTCCAGCACC 298

All pegRNAs for prime editors were of the form (SEQ ID NO: 299) 5′-NNNNNNNNNNNNNNNNNNNNGTTTTAGAGCTAGAAATAGCAAGTTA AAATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCACCGAGTCGGTG CNNNNNNNNNNNNNNNNNNNNTTTTTTT-3′.

TABLE M Shown below are the protospacer and 3′ extension sequences for these pegRNAs (all written 5′ to 3′). target protospacer SEQ ID SEQ ID gene/site sequence NO: 3′ extension sequence NO: ABE_site7_ CTATATTACTT 300 GAATAGTAAGCATAGACTC 301 CtoG ACCTTATCC CAGGATAAGGTAAGTAATAT ABE_site8_ ATGAGGAAAG 302 GTAAAGAAAGCATAGACTG 303 CtoG GGACTAGAGT AGGGGTACAATCCTACTCT AGTCCCTTTCCTC HEK_site2_ GCTGGCCCTG 304 GCTTTCTGTTCCAGTTTCCT 305 CtoG TAAAGGAAAC TTACAGGGCCA RNF2_site1_ TGAGTTACAA 306 GTCATGTTAGTCATTACCTG 307 CtoG CGAACACCTC AGGTGTTCGTTGTAACT HEK_site3_ GGCCCAGACT 308 TCTGCCATCAAAGCGTGCT 309 CTTins GAGCACGTGA CAGTCTG FANCF_site1_ GGAATCCCTT 310 GGAAAAGCGATCAAGGTGC 311 GtoT CTGCAGCACC TGCAGAAGGGA

All nicking gRNAs for PE3 system were of the form  (SEQ ID NO: 145) 5′-NNNNNNNNNNNNNNNNNNNNCGTTTTAGAGCTAGAAATAGCAAGTTAA AATAAGGCTAGTCCGTTATCAACTTGAAAAAGTGGCACCGAGTCGGTGCTT TTTTT-3′. 

TABLE N Shown below are the protospacer regions for these  nicking gRNAs (all written 5′ to 3′). SEQ ID target gene/site PE3 nicking guide RNA NO: ABE_site 7_CtoG AGAATGGCAGGCACGTAGTA 312 ABE_site 8_CtoG TGCAACAGCTATGAAATAGC 313 HEK_site 2_CtoG TTGTTTGCAGCTATTCAGGC 314 RNF2_site 1_CtoG TACACGTCTCATATGCCCCT 315 HEK_site 3 CTTins GTCAACCAGTATCCCGGTGC 316 FANCF_site 1_GtoT GAAGCTCGGAAAAGCGATCA 317

Cell Culture

STR-authenticated HEK293T (CRL-3216), K562 (CCL-243), HeLa (CCL-2), and U2OS cells (similar match to HTB-96; gain of #8 allele at the D5S818 locus) were used in this study. HEK293T and HeLa cells were grown in Dulbecco's Modified Eagle Medium (DMEM, Gibco) with 10% heat-inactivated fetal bovine serum (FBS, Gibco) supplemented with 1% penicillin-streptomycin (Gibco) antibiotic mix. K562 cells were grown in Roswell Park Memorial Institute (RPMI) 1640 Medium (Gibco) with 10% FBS supplemented with 1% Pen-Strep and 1% GlutaMAX (Gibco). U2OS cells were grown in DMEM with 10% FBS supplemented with 1% Pen-Strep and 1% GlutaMAX. Cells were grown at 37° C. in 5% CO2 incubators and periodically passaged upon reaching around 80% confluency. Cell culture media supernatant was tested for mycoplasma contamination using the MycoAlert mycoplasma detection kit (Lonza) and all tests were negative throughout the experiments.

Transfections

HEK293T cells were seeded at 1.25×10⁴ cells per well into 96-well flat bottom cell culture plates (Corning) for DNA on-target experiments or at 6.25×10⁴ cells per well into 24-well cell culture plates (Corning) for DNA off-target experiments. 24 hours post-seeding, cells were transfected with 30 ng of control or base/prime editor plasmid and 10 ng of gRNA plasmid (and 3.3 ng nicking gRNA plasmid for PE3) using 0.3 μL of TransIT-X2 (Mirus) lipofection reagent for experiments in 96-well plates, or 150 ng control or base editor plasmid and 50 ng gRNA, and 1.5 μL TransIT-X2 for experiments in 24-well plates. K562 cells were electroporated using the SF Cell Line Nucleofector X Kit (Lonza), according to the manufacturer's protocol with 2×10⁵ cells per nucleofection and 800 ng control or base/prime editor plasmid, 200 ng gRNA or pegRNA plasmid, and 83 ng nicking gRNA plasmid (for PE3). U2OS cells were electroporated using the SE Cell Line Nucleofector X Kit (Lonza) with 2×10⁵ cells and 800 ng control or base/prime editor plasmid, 200 ng gRNA or pegRNA, and 83 ng nicking gRNA (for PE3). HeLa cells were electroporated using the SE Cell Line 4D-Nucleofector X Kit (Lonza) with 5×10⁵ cells and 800 ng control or base/prime editor, 200 ng gRNA or pegRNA, and 83 ng nicking gRNA (for PE3). 72 hours post-transfection, cells were lysed for extraction of genomic DNA (gDNA).

DNA Extraction

HEK293T cells were washed with 1×PBS (Corning) and lysed overnight by shaking at 55° C. with 43.5 μl of gDNA lysis buffer (100 mM Tris-HCl at pH 8, 200 mM NaCl, 5 mM EDTA, 0.05% SDS) supplemented with 5.25 μl of 20 mg/ml Proteinase K (NEB) and 1.25 μl of 1M DTT (Sigma) per well for experiments in 96-well plates, or with 174 μl DNA lysis buffer, 21 μl Proteinase K, and 5 μL 1M DTT per well for experiments in 24-well plates. K562 cells were centrifuged for 5 min, media removed, and lysed overnight by shaking at 55° C. with 174 μl DNA lysis buffer, 21 μl Proteinase K, and 5 μL 1M DTT per well in 24-well plates. U2OS cells and HeLa cells were washed with 1×PBS and lysed overnight shaking at 55° C. with 174 μl DNA lysis buffer, 21 μl Proteinase K, and 5 μL 1M DTT per well in 24-well plates. Subsequently, gDNA was extracted from lysates using 1-2× paramagnetic beads as previously described⁷ and eluted in 45 μl of 0.1×EB buffer. DNA extraction was performed using a Biomek FX^(P) Laboratory Automation Workstation (Beckman Coulter).

Targeted Amplicon Sequencing

DNA targeted amplicon sequencing was performed as previously described.⁷ Briefly, extracted gDNA was quantified using the Qubit dsDNA HS Assay Kit (Thermo Fisher). Amplicons were constructed in 2 PCR steps. In the first PCR, regions of interest (170-250 bp) were amplified from 5-20 ng of gDNA with primers containing Illumina forward and reverse adapters on both ends (Supplementary Table 9). PCR products were quantified on a Synergy HT microplate reader (BioTek) at 485/528 nm using a Quantifluor dsDNA quantification system (Promega), pooled and cleaned with 0.7× paramagnetic beads, as previously described.⁷ In a second PCR step (barcoding), unique pairs of Illumina-compatible indexes (equivalent to TruSeq CD indexes, formerly known as TruSeq HT) were added to the amplicons. The amplified products were cleaned up with 0.7× paramagnetic beads, quantified with the Quantifluor or Qubit systems, and pooled before sequencing. The final library was sequenced on an Illumina MiSeq machine using the Miseq Reagent Kit v2 (300 cycles, 2×150 bp, paired-end). Demultiplexed FASTQ files were downloaded from BaseSpace (Illumina).

Example 1. ABE Induces C-to-G Editing in Human HEK293T Cells

Human HEK293T cells were transfected with plasmids encoding nCas9, ABEmax, miniABEmax-K20/R21A, and miniABEmax-V82G (FIG. 1-2 ) and gRNAs targeting several genomic sites (e.g. FANCF site 1, HEK site 2 and ABE site 7). After 72 hours, gDNA was extracted and targeted amplicon sequencing was performed to determine the on-target DNA editing of ABE constructs. C-to-G editing was seen on all three sites next to the expectedly robust A-to-G DNA base editing and probably stemmed from deamination of cytosine by the adenosine deaminase TadA, followed by downstream DNA and base excision repair (FIG. 1-4 )

Example 2. Engineering of CGBE1 and its Testing at 25 Genomic Loci

Given the observation outlined in Example 1 on ABE-mediated C-to-G alterations, we wondered whether we could induce these edits more efficiently by modifying the BE4max CBE^(8,15), which harbors an enzyme actually intended to deaminate cytosines (the rat APOBEC1 cytidine deaminase)(FIG. 5-6 ). Removal of the two UGIs from BE4max to create BE4maxΔUGI resulted in an increase in C-to-G (and to a lesser degree C.-to-A) edits relative to wild-type BE4max when tested with seven different gRNAs targeted to sites with Cs at protospacer positions 5, 6, and 7 in HEK293T cells (FIG. 13 ). In general, C-to-G editing was observed with BE4maxΔUGI at Cs that were preceded by A, C, or T, with the most efficient editing generally observed with Cs at protospacer position 6 (FIG. 13 ). We also observed a substantially higher frequency of indels with BE4maxΔUGI relative to BE4max (FIGS. 13 & 15 ), consistent with the idea that this fusion is likely more efficient at creating abasic sites.¹ Reasoning that creation of an abasic site is important for increased C-to-G editing, we further hypothesized that adding human UNG (hUNG) enzyme to BE4maxΔUGI might enhance the frequency of desired edits. However, a BE4maxΔUGI-hUNG fusion possessed somewhat decreased C-to-G editing activity and did not induce appreciably changed frequencies of indels with the seven gRNAs tested (although it did show decreased C-to-T editing activity) (FIGS. 13 & 15 ). Similar results were obtained when hUNG was fused at the N-terminus of BE4maxΔUGI (FIG. 14 ). Fusion of UNG to ABEmax did not yield enhanced C-to-G editing compared to ABEmax (FIG. 14 ). We also tested a variety of CBEs that are based on non-APOBEC1 deaminase architectures, such as human A3A and enhanced A3A-BE3¹⁷, human AID-BE3¹⁵, and the Petromyzon marinus CDA1-based Target-AID², as well as variants thereof lacking UGIs and having added UNGs. Among this larger ensemble of variants, none consistently showed higher activity than the BE4maxΔUGI-hUNG editor (FIG. 16 ).

We also investigated whether introducing mutations into the APOBEC1 part of BE4maxΔUGI-hUNG might further increase the frequency of C-to-G editing. Although we do not have a mechanistic understanding of how C-to-G edits are induced, we reasoned that altering the deamination dynamics of APOBEC1 might also influence the editing outcome. We focused on the APOBEC1 R33A mutation, a substitution we previously showed can decrease off-target RNA editing while substantially preserving the efficiency and increasing the precision of on-target DNA editing by CBEs⁵. We found that introduction of R33A into BE4maxΔUGI-hUNG increased C-to-G editing frequencies with three of the seven gRNAs tested in HEK293T cells while leaving editing frequencies essentially unaltered with the other four (FIG. 13 ). The effect of the R33A variant was most striking with the FANCF site 1 gRNA, which had shown virtually no C-to-G editing with any of the other editors we tested but now showed a mean editing frequency of 14.0% (FIG. 13 ). Interestingly, BE4max(R33A)ΔUGI-hUNG on average showed lower indel byproducts with 6 out of 7 gRNAs compared to BE4maxΔUGI-hUNG (FIG. 15 ).

We additionally explored whether replacing the hUNG present in the BE4max(R33A)ΔUGI editor with an orthologous UNG from Escherichia coli (eUNG) might further increase the efficiency of C-to-G edits. We created two additional editors: BE4max(R33A)ΔUGI-eUNG or eUNG-BE4max(R33A)ΔUGI with an eUNG added to the carboxy- or amino-terminal ends, respectively. Testing of these fusions in HEK293T cells revealed that both induced C-to-G edits with higher frequencies than BE4max(R33A)ΔUGI-hUNG for six out of seven gRNAs tested (mean editing frequencies ranging from 3.3-57.0% and 8.5-62.6% for BE4max(R33A)ΔUGI-eUNG and eUNG-BE4max(R33A)ΔUGI, respectively) (FIG. 13 ). Indel frequencies with both fusions were generally comparable to those observed with BE4max(R33A)ΔUGI-hUNG (FIG. 15 ). Given its higher C-to-G editing activity, we chose the eUNG-BE4max(R33A)ΔUGI fusion (hereafter referred to as C-to-G Base Editor 1 (CGBE1)) for additional characterization.

To more comprehensively characterize CGBE1, we tested its activity with 18 additional gRNAs in human HEK293T cells. 12 of the sites targeted by these 18 gRNAs have a C at position 6 (“C6-sites”) (FIGS. 18 & 20 ) and 6 have a C at positions 4, 5, 7, or 8 (“non-C6-sites”) (FIGS. 18 & 20 ). For 16 of the 18 sites, CGBE1 induced C-to-G edits with substantially higher frequencies than what was observed with its parental CBE control (BE4max(R33A)) (FIG. 18 ). Highly efficient C-to-G edits were observed for 4 of the 18 sites (ABE site 7, ABE site 8, HEK site 2, and PPP1R12C site 6), with mean editing frequencies ranging from 41.7 to 71.5% (FIG. 18 ). C-to-G edits were by far the most efficiently induced edits at these 4 sites with only very low levels of C-to-T or C-to-A byproducts observed (FIG. 18 ). C-to-G was also the most efficiently induced edit for 6 additional sites albeit at lower frequencies (three C6-sites and three non-C6-sites) (FIG. 18 ). In total, when combined with the results obtained with the initial seven gRNAs described above (FIG. 13 ), CGBE1 induced C-to-G editing with mean frequencies of 20% or higher at 14 of the 25 sites tested (FIGS. 13 & 18 ). Notably, C-to-G editing was most efficient for Cs embedded in an AT-rich sequence context (FIGS. 13 & 18 ). Analysis of the spatial distribution of editing across all 25 sites tested shows that the mean frequency of C-to-G editing was highest at position 6 and that indels were distributed throughout the length of the protospacer (FIG. 19 ).

Example 3. Characterization of miniCGBE1 and its Side by Side Comparison with CGBE1

We explored the impact of deleting the eUNG domain from the CGBE1 editor on its activity. This particular editor architecture, which we named miniCGBE1 (FIG. 22 ), had not been made or tested over the course of the stepwise progression from BE4max to CGBE1 and also has the added advantage of being smaller in size. Side-by-side comparisons of miniCGBE1 with CGBE1 at the same 25 sites we had previously tested showed that the frequencies of editing observed with miniCGBE1 were comparable but moderately lower at 6 out of 25 sites tested (mean editing frequencies across all 25 sites of 14.4% and 13% with CGBE1 and miniCGBE1, respectively), whereas the indel frequencies induced by miniCGBE1 were lower at 15 out of 25 sites (mean indel frequencies of 10.4% and 8.5% for CGBE1 and miniCGBE1, respectively; FIG. 22-24 ).

To more fully characterize the positional preferences within the editing windows of CGBE1 and miniCBGE1, we tested these two editors side-by-side with BE4max and BE4max(R33A) using 23 additional gRNAs that target sites with cytosines at protospacer positions 4, 5, 7, and 8 (FIG. 25 ). The targets of these 23 gRNAs included six sites with a C5, five with a C7, four with a C8, and eight with two Cs at various positions (C4 and C7, C4 and C8, C5 and C7, C5 and C8, and C7 and C8). Mean editing frequencies induced by miniCGBE1 were comparable to those of CGBE1: 1.7% and 1.5% at C4, 7.3% and 6.7% at C5, 16.0% and 13.5% at C7 and 3.4% and 2.9% at C8 for CGBE1 and miniCGBE1, respectively (FIG. 25 ). In addition, indel frequencies induced by CGBE1 and miniCGBE1 were comparable at 10 sites, lower with CGBE1 at five sites, and lower with miniCGBE1 at eight sites (FIG. 26 ). Collectively, our testing of CGBE1 and miniCGBE1 with 48 different gRNAs demonstrates that both have an optimal editing window for cytosines at positions 5-7 in the protospacer with those at position 6 being edited most efficiently (FIG. 27 ). This finding is consistent with our previously published studies showing that a CBE with the APOBEC1-R33A variant edits optimally on positions 5-7 of the protospacer and more weakly on positions 4 and 87.

Example 4. Evaluation of DNA Off-Target Editing Induced by CGBE

Cas9-dependent DNA off-target profiles of CGBEs was assessed by transfecting HEK 293T cells with nCas9 control, BE4max, BE4max(R33A), CGBE1, and miniCGBE1 using HEK site 2, HEK site 3, HEK site 4, EMX1 site 1, and FANCF site 1 gRNAs. 23 genomic sites that have previously been described as known off-target sites for said gRNAs (Tsai et al, NBT 2014) were sequenced with NGS to detect potential off-target base editing of CGBE constructs. BE4max induced C-to-D (D=A, G, or T) edits at 15 of the 23 off-target sites with BE4max-R33A inducing edits less efficiently at all 15 sites, consistent with previously published observations that introduction of R33A reduces Cas9-dependent DNA off-target edits by the BE3 CBE (FIG. 28 ). Similarly, both CGBE1 and miniCGBE1 showed lower C-to-D off-target editing at 14 out of the 15 off-target sites that were edited by BE4max (FIG. 28 ). As expected, off-target indel frequencies were higher with CGBE1 and miniCGBE1 relative to BE4max at 18 out of 23 sites, although miniCGBE1 again showed reduced activity compared with CGBE1 at 14 out of these 18 sites (FIG. 28 ). Overall, this assessment of Cas9/gRNA-dependent DNA off-target editing shows that CGBE1 and miniCGBE1 induce fewer off-target DNA edits than BE4max, that CGBE-induced indels can occur at off-target sites, and that indels are reduced with miniCGBE1 relative to CGBE1.

Example 5. CGBEs with SpCas9-NG and SpCas9-VRQR Variants of are Functional

We tested whether we could improve the somewhat more restricted targeting range of CGBEs by using previously described SpCas9-NG and SpCas9-VRQR variants that recognize shorter NG¹⁹ and alternative NGA²⁰ PAMs, respectively. We targeted six sites with NGT PAMs using modified CGBE1-NG and miniCGBE1-NG variants and six sites with NGAG PAMs using CGBE1-VRQR and miniCGBE1-VRQR variants. Each of these 12 new sites have a cytosine at position 6 embedded within an AT-rich sequence context to provide an optimal target for C-to-G editing (FIG. 30 ). On these target sites, CGBE1-NG and miniCGBE1-NG induced C-to-G edits with frequencies as high as 27% and 26%, respectively, and CGBE1-VRQR and miniCGBE1-VRQR induced C-to-G edits with frequencies of up to 31% (FIG. 30 ). These results show that the targeting range of CGBE constructs can be expanded by using Cas9 variants with altered or relaxed PAM recognition specificities.

Example 6. Comparison of CGBEs with Prime Editor Technologies

We compared our CGBEs with Prime Editing (PE) methods that can introduce a diverse range of different edits and that were published15 while we were completing this project. The PE2 system uses two components: (1) a Prime Editor fusion protein and (2) a prime editing gRNA (pegRNA) (FIG. 32 ).²¹A more efficient PE3 system adds a secondary “nicking gRNA” (ngRNA) that directs a nick to the DNA strand opposite the edited one, thereby increasing editing efficiency (FIG. 32 ).²¹ We performed side-by-side comparisons of our CGBEs with PE2 and PE3 systems for making four different C-to-G edits, assessing frequencies of these alterations across four different human cell lines (HEK293T, K562, U205, and HeLa cells). Positive control experiments we performed in all four cell lines re-confirmed that two other previously described pegRNAs could induce a G-to-T transversion in FANCF site 1 and a CTT insertion in HEK site 3 (CTT-insertion), that PE3 is more efficient than PE2, and that the highest prime edit frequencies were observed in HEK293T cells (FIG. 33 ). For all four C-to-G edits (which we had already established could be efficiently induced by our CGBEs in HEK293T cells), we found that both PE2 and PE3 were substantially less efficient than CGBE and miniCGBE1 across all four cell lines (FIGS. 34 & 36 ). Importantly, these data also establish that our CGBEs can function robustly and efficiently across multiple human cancer cell lines. In addition, we found that the frequencies of unwanted indels were lower with prime editors compared to the CGBEs in all four cell lines (FIG. 37 ). To rule out that the pegRNAs and ngRNAs we designed were inactive or unable to interact with Cas9, we tested their abilities to induce Cas9-mediated indels at their intended target sites in HEK293T cells (note that we could not assess the activity of the HEK site 3 ngRNA due to its overlap with a required PCR primer). The indel frequencies we observed with these pegRNAs and ngRNAs were comparable to those used with the two positive control target sites (FIG. 35 ).

Example 7. mRNA and RNP Production of CGBEs and Testing in Primary Human CD34+ and T Cells

CGBE architectures described in FIGS. 6-9 will be tested in primary human CD34+ and T cells by electroporating CGBE mRNAs (produced via IVT or by TriLink). CGBE constructs will be subcloned into pET vectors with an N-terminal 6×His-tag and codon-optimized for expression in E. coli to enable protein purification. RNPs will be electroporated with a Lonza device into HEK293T and primary human T cells to determine if CGBE RNP delivery yields efficient ex vivo DNA transversion base editing.

Example 8. Evaluation of RNA Off-Target Editing Induced by CGBE

Unbiased detection of RNA off-target editing with the help of RNA-seq will be assessed. Cells will be transfected with two different gRNAs and CGBE constructs that are co-translationally expressed with P2A-EGFP in 15 cm dishes and trypsinized 36 hours post-transfection. Subsequently, GFP+ cells will be sorted on a BD FACSAria II and lysed to harvest both DNA and RNA. After efficient on-target editing is confirmed via targeted amplicon sequencing, RNA-seq will be performed using a TruSeq stranded total RNA library prep and sequencing on a NextSeq 500 machine at the MGH or a NovaSeq at the Broad Institute.

Example 9. Evaluation of UNG Recruitment Strategy Using Peptide Aptamers

Next generation CGBE constructs fused with the candidate peptide aptamers will be assessed by transfection experiments, for example, those using lipofection and nucleofection techniques into human cells such as HEK 293T, U2OS and K562 cell lines. The transfections will be carried out with gRNA constructs with spacer sequences targeting human genomic loci having cytosines in the editing windows that is generated by our CGBE constructs. 72 hours post-transfection, genomic DNA (gDNA) will be harvested, and target loci will be PCR amplified. PCR amplicons will be subjected to targeted next generation sequencing (NGS) to quantify on-target editing efficiencies. The DNA off-target activities of the next generation CGBE constructs will be assessed by analyzing the top in-silico predicted candidate off-target sites using targeted amplicon sequencing (NGS) using the treated gDNAs. In order to assess the potential RNA off-target activities of our next generation CGBE constructs, we will be harvesting total RNA in parallel in the treated cells in order to conduct stranded libraries for transcriptome-wide analysis via RNA sequencing (RNA-seq).

Example 10. Evaluation of UNG Recruitment Strategy Using RNA Aptamers

The next generation CGBE constructs will be analyzed using RNA aptamers fused to the gRNA in a series of transfection experiments (using, for example, lipofection and nucleofection techniques) in human cells such as HEK 293T, U2OS and K562 cell lines. The transfections will be carried out with fusion gRNA constructs with spacer sequences targeting human genomic loci having cytosines in the editing windows generated by our CGBE constructs. 72 hours post-transfection, genomic DNA (gDNA) will be harvested, and target loci will be PCR amplified. PCR amplicons will be subjected to targeted next generation sequencing (NGS) to quantify on-target editing efficiencies. In order to test the potential DNA off-target activities of our next generation CGBE constructs, the top in-silico predicted candidate off-target sites will be analyzed with targeted amplicon sequencing (NGS) using the treated gDNAs. In order to assess the potential RNA off-target activities of our next generation CGBE constructs, we will be harvesting total RNAs in parallel in the treated cells in order to conduct transcriptome-wide analysis via RNA sequencing (RNA-seq).

Example 11. Evaluation of UNG Recruitment Strategy Using Fabs, scFVs or sdABs

Next generation CGBE constructs fused with the candidate Fab, scFv, or sdAb, will be assessed in a series of transfection experiments (e.g., using lipofection or nucleofection techniques) in human cells such as HEK 293T, U2OS and K562 cell lines. The transfections will be carried out with gRNA constructs with spacer sequences targeting human genomic loci having cytosines in the editing windows generated by CGBE constructs. 72 hours post-transfection, genomic DNA (gDNA) will be harvested, and target loci will be PCR amplified. PCR amplicons will be subjected to targeted next generation sequencing (NGS) to quantify on-target editing efficiencies. DNA off-target activities of the next generation CGBE constructs will be assessed by analyzing the top in silico predicted candidate off target sites using targeted amplicon sequencing (NGS). In order to assess the potential RNA off-target activities of our next generation CGBE constructs, we will be harvesting total RNA in parallel in the treated cells in order to conduct transcriptome-wide analysis via RNA sequencing (RNA-seq).

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EXEMPLARY SEQUENCES SEQ ID NO: 1 >tr|G3U0R4|G3U0R4_LOXAF Apolipoprotein B mRNA editing enzyme catalytic subunit 1 OS = Loxodonta africana = african elephant OX = 9785 GN = APOBEC1 PE = 4 SV = 1 FRRRIKPWEFEIFFDPRQLRKETCLLYEIKWGTSHKVWRNSGQNTTKHVEVNFIEKFTSERK LCPSISCSITWFLSWSPCWECSKAIREFLRQHPNVTLVIYVARLFHHMDQRNRQGLKDLILS GITVQIMRVSEYHHCWRNFVSYSPGEETYWPRYPPLWMMMYALELHCIILSLPPCLKISRR CQHQLTLFSLTPQKCHYQMIPPYILLATGLIEPPMTWR SEQ ID NO: 2 >tr|A0A0M3N0G8|A0A0M3N0G8_PROAN APOBEC-1 OS = Protopterus annectens OX = 7888 PE = 2 SV = 1 MVQKRTSASKTRMTKKVLLSEYQKFYYSPRTCIGYVIQYDEDNVIFQNWICNKRTTHAELQC IYEIKQNSLIKRFTPCTLKWYMSWTPCSECANEIIRFLNKFCQVKLEICAARIYFHKK KDNRRALRNLVKAGVKLTTMRWKDYKSMWRRFGTGEEIKKYEFFEKSSDHKSVNWRWTL KKILKEKDRDSDLENALSLLKI SEQ ID NO: 3 >tr|A0A151P6M4|A0A151P6M4_ALLMI C->U-editing enzyme APOBEC-1 OS = Alligator mississippiensis OX = 8496 GN = APOBEC1A PE = 4 SV = 1 MAVEEEKGLLGTSQGWKIELKDFQENYMPSTWPKVTHLLYEIRWGKGSKVWRNWCSNTL TQHAEVNCLENAFGKLQFNPPVPCHITWFLSWSPCCQCCRRILQFLRAHSHITLVIKAAQLF KHMDERNRQGLRDLVQSGVHVQVMDLPDYRYCWRTFVSHPHEGEGDFWPWFFPLWITF YTLELQHILLQQHALSYNL SEQ ID NO: 4 >tr|F1CGT0|F1CGT0_ANOCA Apolipoprotein B mRNA-editing enzyme 1a isoform (Fragment) OS = Anolis carolinensis OX = 28377 PE = 2 SV = 1 KAAILLSNLFFRWQMEPEAFQRNFDPREFPECTLLLYEIHWDNNTSRNWCTNKPGLHAEEN FLQIFNEKIDIKQDTPCSITWFLSWSPCYPCSQAIIKFLEAHPNVSLEIKAARLYMHQI DCNKEGLRNLGRNRVSIMNLPDYRHCWTTFVVPRGANEDYWPQDFLPAITNYSRELDSILQ D SEQ ID NO: 5 >tr|A0A091EQ78|A0A091EQ78_CORBR C->U-editing enzyme APOBEC-1 (Fragment) OS = Corvus brachyrhynchos OX = 85066 GN = N302_10757 PE = 4 SV = 1 RWKIEPGDFQINYSPSQHRRGVYLLYEIRWRRGSIWRNWCSNTHRQHAEVNFLENCFKDR PQVPCSITWFLSASPCGKCSKRILEFLKSRPYVTLKIYAAKLFRHHDIRNREGLCNLGMHGV TIHIMNLEDYSYCWRNFVVY SEQ ID NO: 6 >tr|A0A091IIG0|A0A091IIG0_CALAN C->U-editing enzyme APOBEC-1 (Fragment) OS = Calypte anna OX = 9244 GN = N300_12023 PE = 4 SV = 1 RWKIQPNDFKRNYQPGRRPNWYLLYEIRWRRGTIWRNWCSNEFPQHAEDNFFQNRFNA VPSVSCSITWFLSTTPCGRCSKRILEFLRLHPNVTLKIYAARLFRHLDNRNRQGLRKLASNG VIIQIMGLPDYSYSWKKFVAY SEQ ID NO: 7 >tr|A0A2U4ALA1|A0A2U4ALA1_TURTR C->U-editing enzyme APOBEC-1 isoform X1 OS = Tursiops truncatus OX = 9739 GN = APOBEC1 PE = 4 SV = 1 MIICWSTGPSAGDATLRRRIEPWEFEVSFDPRELSKETRLLYEIKWGKSQRIWRHSGKNTT KHVERNFIEQITSERRFHRSVSCCIIWFLSWSPCWECSEAIREFLKQHPRVTLLIYVARLFQH MDPRNRQGLRDLTHSGVTIQIMGPTEYDYCWRYFVNYAPGKEAHWPRYPPLLMKLYALEL HCIILGLPPCLNISRYQNQLTLFRPILRNCHYQMIPPHILLHTGLIQLPLTWR SEQ ID NO: 8 >tr|A0A093FY71|A0A093FY71_TYTAL C->U-editing enzyme APOBEC-1 (Fragment) OS = Tyto alba 0X = 56313 GN = N341_11998 PE = 4 SV = 1 RWKIQPNDFKRNFLPGQHPKVVYLMYEIRWIRGTAWRSWCSNNSKQDAEVNLLENCFKA MPSVFCSVTWVLFTTPCGKCFRRILEFLRVHSNVALERYAAQLFRHLDICNWQGIRSLAMN GVIIHIMNLADYSYCWKRFVAY SEQ ID NO: 9 >tr|L5KGJ8|L5KGJ8_PTEAL C->U-editing enzyme APOBEC-1 OS = Pteropus alecto OX = 9402 GN = PAL_GLEAN10015600 PE = 4 SV = 1 MWVLFDILISWSTGPSTGDPTLRRRIEPWEFEVFFDPRELRKEACLLYEIQWGTSHKIWRNS GKNTTKHVELNFIEKFTSERHFCSSVSCSIIWFLSWSPCWECSKAIREFLSQRPTVTLVIFVS RLFQHMDQQNRQGLRDLINSGVTIQIMRASEYDHCWRNFVNYPPGKEAHWPRYPPLWMK LYALELHCIILSLPPCVMISRRCQKQLTLFTLILKKCHYQMIPAHILLATGLIQVPVTWR SEQ ID NO: 10 >tr|A0A2K6KS69|A0A2K6KS69_RHIBE CMP/dCMP-type deaminase domain-containing protein OS = Rhinopithecus bieti OX = 61621 GN = APOBEC1 PE = 4 SV = 1 MTSEKGPSTGDPTLRRRIEPWEFDIFYDPRELRKEACLLYEIKWGMSWKIWRSSGKNTTNH VEVNFIEKFTSERRFHSSISCSITWFLSWSPCWDCSQAIRKFLSQHPGVTLVIYVARLFWHT DQQNRQGLRDLVNSGVTIQMMTASEYYHCWRNFVNYPPGEEAHWPRYPPLWMMLYALE LHCIILSLPPCLKISRRWQNHLTFFRLHLQNCHYQTIPPHILLATGLIQPSVTWR SEQ ID NO: 11 >tr|A0A2Y9NGP5|A0A2Y9NGP5_DELLE C->U-editing enzyme APOBEC-1 isoform X1 OS = Delphinapterus leucas OX = 9749 GN = APOBEC1 PE = 4 SV = 1 MIICWSTGPSAGDATSRRRIEPWEFEVSFDPRELCKETRLLYEIKWGKSQHVWRHSDKNTT KHVECKFIEKITSERHFHPSVSCCIIWFLSWSPCWECSKAIREFLNQHPRVTLFIYVARLFQH MDPQNRQGLRDLIHSGVTIHVMGPTEYDYCWRNFVNYPPGKEAHWPRYPPMLMKLYALE LHCIILGLPPCLNISRYQNQLTLFRLIPQNCHYRMIPPHILLHRGLIRLPLTWR SEQ ID NO: 12 >tr|A0A218ULD2|A0A218ULD2_9PASE C->U-editing enzyme APOBEC-1 OS = Lonchura striata domestica OX = 299123 GN = APOBEC1 PE = 4 SV = 1 MYRRKMRGMYISKRALRKHFDPRNYPRETYLLCELQWRGSHKSWQHWLRNDDSKDCHA EKYFLEEIFEPRSYNICDMTWYLSWSPCGECCDIIQDFLEEQPNVNINIRIARLYYADRASNR RGLMELANSPGVSIEIMDADDYNDCWETFIQPGVYYRFSPENFESAIRRNCSQLEDILQGLH L   SEQ ID NO: 13 >tr|A0A0Q3WRD0|A0A0Q3WRD0_AMAAE C->U-editing enzyme APOBEC-1 OS = Amazona aestiva OX = 12930 GN = AAES_27783 PE = 4 SV = 1 MLPAPAPVPLVLPLQGGGVVVVTVGVXPTALLQPSGAPEVARTFVGAVIAFVIAEYVDTSVS EDTTICGMYIPKEALKYHFDPREVXRDTYLLCILRWGETGTPWSHWVKNYRYHAEVYFLEKI FQTRKSSKNINCSITWYLSWSPCAKCCRKILNFLKKHSYVSIKIHVARLFRIDDKETXQNLKN LGSLVGVTVSVMEXEDYTNCWKTFIRGHADGDSWIDDLKSEIRKNRLKFQGIFKDLPHQTE DVDFWLILAANPGPAWFSFSGYTGWAVASKAPSLLSPLSCLTRLLTP SEQ ID NO: 14 >tr|A0A2K6U925|A0A2K6U925_SAIBB Apolipoprotein B mRNA editing enzyme catalytic subunit 1 OS = Saimiri boliviensis boliviensis OX = 39432 GN = APOBEC1 PE = 4 SV = 1 MTSERRRIEPWEFSISYDPRELCKETCLLYEIKWGMSWKIWRSSGKNTTNHVEVNFIEKFTS ERHFHSSVSCSITWFLSWSPCWECSQAIREFLSQHPGVTLVIYVARLFQHMDQQNRQGLR ELVNSGVTIQIMTASEYYHCWRNFVNYPPGEEAHWPRHPPLWMMLYALELHCIIL SEQ ID NO: 15 >tr|A0A2R9A0R0|A0A2R9A0R0_PANPA CMP/dCMP-type deaminase domain-containing protein OS = Pan paniscus OX = 9597 GN = APOBEC1 PE = 4 SV = 1 ISWSTGPSTGDPTLRRRIEPWEFDVFYDPRELRKEACLLYEIKWGMSRKIWRSSERDFHPSI SCSITWFLSWSPCWECSQAIREFLSQHPGVTLVIYVARLFWHMDQQNRQGLRDLVNSGVTI QIMTASEYYHCWRNFVNYPPGDEAHWPQYPPLWMMLYALELHCIILSLPPCLKISRRWQN HLTFFSLHLQNCHYQTIPPHILLATGLIHPSVAWR SEQ ID NO: 16 >sp|Q694B3|ABEC1_PONPY C->U-editing enzyme APOBEC-1 OS = Pongo I OX = 9600 GN = APOBEC1 PE = 3 SV = 2 MTSEKGPSTGDPTLRRRIESWEFDVFYDPRELRKETCLLYEIKWGMSRKIWRSSGKNTTNH VEVNFIKKFTSERRFHSSISCSITWFLSWSPCWECSQAIREFLSQHPGVTLVIYVARLFWHM DQRNRQGLRDLVNSGVTIQIMRASEYYHCWRNFVNYPPGDEAHWPQYPPLWMMLYALEL HCIILSLPPCLKISRRWQNHLAFFRLHLQNCHYQTIPPHILLATGLIHPSVTWR SEQ ID NO: 17 >tr|E1BP99|E1BP99_BOVIN Apolipoprotein B mRNA editing enzyme catalytic subunit 1 OS = Bos taurus OX = 9913 GN = APOBEC1 PE = 4 SV = 1 MASDRGPPAGDPTLRRRIEPWEFEFSFDPRKFCKEACLLYEIQWGNNRDVWRHSGKNTTK HVERNFIEKIASERYFCPSIRCFIFWYLSWSPCWECSKAIREFLNQHPNVTLVIYIARLFQHM DPQNRQGLKDLVQSGVTIQVMRAPEYEYCWRNFVNYPRGKEAHWPRYPPLWMNLYALEL YCIILGLPPCLHISRRYQNQLIVFRLTLQNCHYQMIPPYILLATGMVQLPMTWR SEQ ID NO: 18 >tr|S7PYX0|S7PYX0_MYOBR C->U-editing enzyme APOBEC-1 OS = Myotis brandtii OX = 109478 GN = D623_10002956 PE = 4 SV = 1 MDEQNRQGLRDLIKSGVTVQIMTTPEYDYCWRNFVNYPPGKDTHCPMYPPLWMKLYALEL HCIILSLPPCLMISRRCQKQLTWYRLNLQNCHYQQIPHHILLATVWI SEQ ID NO: 19 >tr|M3WB96|M3WB96_FELCA Apolipoprotein B mRNA editing enzyme catalytic subunit 1 OS = Felis catus OX = 9685 GN = APOBEC1 PE = 4 SV = 2 MASDKGPSAGDATLRRRIEPREFEVFFDPRELRKEACLLYEIKWGTSHRIWRNSGRNTANH VELNFIEKFTSERHFCPSVSCSITWFLSWSPCWECSKAIRGFLSQHPSVTLVIYVSRLFWHL DQQNRQGLRDLVNSGVTVQIMRVPEYDHCWRNFVNYPPGEEDHWPRYPVVWMKLYALE LHCIILSLPPCLKILRRCQNQLTLFRLTLQNCHYQMIPPHILLATGLIQLPVTWR SEQ ID NO: 20 >tr|A0A2K5PZC0|A0A2K5PZC0_CEBCA CMP/dCMP-type deaminase domain-containing protein OS = Cebus capucinus imitator OX = 1737458 GN = APOBEC1 PE = 4 SV = 1 MTSERGPSTGDPTLRRRIEPWEFYISYDPKELCKETCLLYEIKWGMSWKIWRSSGKNTTNH VEVNFIEKFTSERRFHSSISCSITWFLSWSPCWECSQAIREFLSQHPGVTLVIYVARLFQHM DQQNRQGLRDLVNSGVTIQIMRASEYYYCWRNFVNYPPGEEAHWPRHPPLWMMLYALEL HCIILGLPPCLKISRRRQNRLTFFRLHLQNCHYQMIPPHILLAAGLIQPSVTWR SEQ ID NO: 21 >tr|H2Q5C6|H2Q5C6_PANTR Apolipoprotein B mRNA editing enzyme catalytic subunit 1 OS = Pan troglodytes OX = 9598 GN = APOBEC1 PE = 4 SV = 1 MTSEKGPSTGDPTLRRRIEPWEFDVFYDPRELRKEACLLYEIKWGMSRKIWRSSGKNTTN HVEVNFIKKFTSERHFHPSISCSITWFLSWSPCWECSQAIREFLSQHPGVTLVIYVARLFWH MDQQNRQGLRDLVNSGVTIQIMRASEYYHCWRNFVNYPPGDEAHWPQYPPLWMMLYAL ELHCIILSLPPCLKISRRWQNHLTFFSLHLQNCHYQTIPPHILLATGLIHPSVAWR SEQ ID NO: 22 >tr|A0A1U7S7K7|A0A1U7S7K7_ALLSI C->U-editing enzyme APOBEC-1-like OS = Alligator sinensis OX = 38654 GN = LOC102373005 PE = 4 SV = 1 MGEHWQYAGSGEYIPQDQFEENFDPSVLLAETHLLSELTWGGRPYKHWYENTEHCHAEIH FLENFSSKNRSCTITWYLSWSPCAECSARIADFMQENTNVKLNIHVARLYLHDDEHTRQGL RYLMKMKRVTIQVMTIPDYTYCWNTFLEDDGEDESDDYGGYAGVHEDEDESDDDDYLPTH FAPWIMLYSLELSCILQGFAPCLKIIQGNHMSPTFQLHVQDQEQKRLLEPANPWGAD SEQ ID NO: 23 >tr|G3HS7|G3HS7_CRIGR C->U-editing enzyme APOBEC-1 OS = Cricetulus griseus OX = 10029 GN = I79_017346 PE = 4 SV = 1 MTEQEYCYCWRNFVNYPPSNEVYWPRYPNVWMRMYALELYCIVLGLPPCLKIIRRHQHPL TFFTLHLQSCHYQRIPPHILWATGLV SEQ ID NO: 24 >tr|A0A094MFH1|A0A094MFH1_ANTCR C->U-editing enzyme APOBEC-1 (Fragment) OS = Antrostomus carolinensis OX = 279965 GN = N321_09417 PE = 4 SV = 1 RWKMQPNDFKRNYLPVQYPNMVYLLYEIRWSTGTIWRNWCSNNSTQHAEVNFLENRFNS RPSVSCSITWVLSTTPCGKCSTKILEFLRLHPNVTLKIYAAKLFKHLDIRNRQGLRNLAMNGV IIRIMNLADYSYCWKTFVAY SEQ ID NO: 25 >tr|A0A2K6EVT9|A0A2K6EVT9_PROCO CMP/dCMP-type deaminase domain-containing protein OS = Propithecus I OX = 379532 GN = APOBEC1 PE = 4 SV = 1 MTSEKRRIEPWEFEAFFDPRELRKEACLLYEIKWGASHKIWRNTGKSTTRHVEVNFIEKFTS ERRSDSLISCSITWFLSWSPCWECSKAIREFLSQHPNVTLVIYVARLFWHMNQQNRQGLRD LINSGVTVQIMGVSEYCHCWRNFVNYPPGKEASCPTYPPLWMTLYALELHCIILSLPPCLKIS RRCQNQLTFFRLTPQNCHYQTIPPHILLATGLIQPSVTWR SEQ ID NO: 26 >tr|G8F4P7|G8F4P7_MACFA C->U-editing enzyme APOBEC-1 (Fragment) OS = Macaca fascicularis OX = 9541 GN = EGM_20518 PE = 4 SV = 1 GPSTGDPTLRRRIEPWEFDIFYDPRELRKEACLLYEIKWGMSPKIWRSSGKNTTNHVEVNFI EKLTSERRFHSSISCSITWFLSWSPCWECSQAIREFLSQHPGVTLVIYVARLFWHTDQQNR QGLRDLVNSGVTIQIMRASEYYHCWRNFVNYPPGEEAHWPRYPPLWMMLYALELHCIILSL PPCLKISRRWQNHLTFFRLHLQNCHYQMIPPHILLATGLIQPSVTWR SEQ ID NO: 27 >tr|A0A091V7F8|A0A091V7F8_NIPNI C->U-editing enzyme APOBEC-1 (Fragment) OS = Nipponia nippon OX = 128390 GN = Y956_13652 PE = 4 SV = 1 RWKIQPNDFRSNYLPCQHPRVVYLLYEIRWSRGTIWRNWCSNNSTQHAEVNFLENCFKAM PSVPCSITWVLSTTPCGKCSRRILEFLRVHPNVTLEIYAAKLFKHLDIRNRQGLRNLAKNGVV IRIMKLADYSYWWKRFVAY SEQ ID NO: 28 >tr|A0A091SSF0|A0A091SSF0_PELCR C->U-editing enzyme APOBEC-1 (Fragment) OS = Pelecanus crispus OX = 36300 GN = N334_11718 PE = 4 SV = 1 RWKLQPEDFKRNYLPGQHPKVVYLLYEIRWSRGTIWRSWCSNNSKQHAEVNFLENCFKAR PSVSCSITWVLSTTPCGKCSRRILEFLRVHPNVTLEIYAAKLFKHLDIRNQQGLRNLAMNGVII RIMNLADYSYCWKRFVAH SEQ ID NO: 29 >tr|A0A091CVE5|A0A091CVE5_FUKDA C->U-editing enzyme APOBEC-1 OS = Fukomys damarensis OX = 885580 GN = H920_16562 PE = 4 SV = 1 MSDPEFCHCWRNFVNYPPGQEARWPRFPPVWTMLYTLELCCVLLNLPPCLKISRRCHNQL AFFQLNLQNCHYRAIPPAVLFAVGLIHPFVAWA SEQ ID NO: 30 >tr|L5LUG3|L5LUG3_MYODS C->U-editing enzyme APOBEC-1 OS = Myotis davidii OX = 225400 GN = MDA_GLEAN10003736 PE = 4 SV = 1 MASDAGKMDRGPVSFIVLKSVETLCVRRIEPWEFEAIFDPRELRKEACLLYEIKWGTGHKIW RHSGKNTTRHVEVNFIEKITSERQFCSSTSCSIIWFLSWSPCWECSKAITEFLRQRPGVTLVI YVARLYHHMDEQNRQGLRDLVKSGVTVQIMTTPEYDYCWRNFVNYPPGKDTHCPIYPPLL MKLYALELHCIILSLPPCLMISRRCQKQLTWYRLNLQNCHYQQIPHHILLATAWI SEQ ID NO: 31 >tr|F1PUJ5|F1PUJ5_CANLF Apolipoprotein B mRNA editing enzyme catalytic subunit 1 OS = Canis lupus familiaris OX = 9615 GN = APOBEC1 PE = 4 SV = 2 MASDKGPSAGDATLRRRIEPWEFEGFFDPRELRKETCLLYEIQWGTSHKTWRNSGKNTTN HVEINFMEKFAAERQYCPSIRCSITWFLSWSPCWECSNAIRGFLSQHPSVTLVIYVARLFWH TDPQNRQGLRDLINSGVTIQIMTVPEYDHCWRNFVNYPPGKEDHWPRYPVLWMKLYALEL HCIILNLPPCLKISRRNQHQLTLFRLTLQDCHYQTIPPPILLDMGLIQPLVTWR SEQ ID NO: 32 >tr|A0A093GVH6|A0A093GVH6_DRYPU C->U-editing enzyme APOBEC-1 (Fragment) OS = Dryobates pubescens OX = 118200 GN = N307_04563 PE = 4 SV = 1 RWKIHPDEFKLNYVPVGRPRWYLLYEIRWSRGSIWRNWCSNSSTQHAEVNFLENCFKAM PSVSCSITWFLSTTPCGNCSRRILEFLRAHPKVTLAIHAAKLFKHLDVRNRHGLKALATDGVV LHIMSIADYRYCWTKFVAY SEQ ID NO: 33 >tr|A0A2K5Z8Y4|A0A2K5Z8Y4_MANLE CMP/dCMP-type deaminase domain-containing protein OS = Mandrillus leucophaeus OX = 9568 GN = APOBEC1 PE = 4 SV = 1 MTSEKGPSTGDPTLRRRIEPWEFDIFYDPRELRKEACLLYEIKWGMSPKIWRSSGKNTTNH VEVNFIEKLTSERRFHSSISCSITWFLSWSPCWECSQAIREFLSQHPGVTLVIYVARLFWHT DQQNRQGLRDLVNSGVTIQIMRASEYYHCWRNFVNYPPGEEAHWPRYPPLWMMLYALEL HCIILSLPPCLKISRRQQNHLTFFRLHLQNCHYQTIPPHILLATGLIQPSVTWR SEQ ID NO: 34 >tr|A0A087VMP5|A0A087VMP5_BALRE C->U-editing enzyme APOBEC-1 (Fragment) OS = Balearica regulorum gibbericeps OX = 100784 GN = N312_10691 PE = 4 SV = 1 RWKIQPDDFKRNYLPGKHPRWYLLYEIRWSRGTIWRSWCSNNATQHAEINFLETCFLART SVSCSITWVLSTTPCGKCSRRILEFLNAYPNVTLEIYAAKLFRHLDNRNRQGLRNLAMKGVR IHIMNLADYSYFWKIFVAY SEQ ID NO: 35 >tr|A0A087QNJ5|A0A087QNJ5_APTFO C->U-editing enzyme APOBEC-1 (Fragment) OS = Aptenodytes forsteri OX = 9233 GN = AS27_08049 PE = 4 SV = 1 RWKIRPNDFKRNYLPGQHPKVVYLLYEIRWSRGTIWRNWCSNNSTQHAEVNFLENCFKAM PSVSCSITWVLSTTPCGKCSRRILEFLRVHPNVTLEIYAAKLFKHLDIRNRQGLRNLAMNGVII RIMNLADYSYGWKRFVAY SEQ ID NO: 36 >tr|A0A2Y9IYV0|A0A2Y9IYV0_ENHLU C->U-editing enzyme APOBEC-1 OS = Enhydra lutris kenyoni OX = 391180 GN = LOC111142361 PE = 4SV = 1 MASDKGPSAGDATLRRRIEPWEFEVFFDPRELRKEACLLYEIQWGTSHKMWRNTGKNTAN HVELNFIEKFTSERRYCPSTHCSITWFLSWSPCWECCKAIRGFLSQHPSVTLVIYVTRLFWH MDPQNRQGLRDLLKSGVTVQIMRAPEYDHCWKNFVNYPPGKEDHWPRYPELWMKLYELE LYCIILSLPPCLKISRRNQNQLTLFRLTLQNCHYQIIPPHILLDTGLIQLPVIWR SEQ ID NO: 37 >tr|B2NIW5|B2NIW5_MUSPF Apolipoprotein B mRNA editing protein OS = Mustela putorius furo OX = 9669 GN = APOBEC1 PE = 2 SV = 1 MASDKGPSAGDATLRRRIEPWEFEVFFDPRELRKEACLLYEIQWGTSHKMWRNTGKNTAN HVELNFIEKFTSERRYCPSTHCSITWFLSWSPCWECSKAIRGFLSQCPSVTLVIYVTRLFWH MDPQNRQGLRDLLKSGVTVRIMRAPEYDHCWKNFVNYPPGKEDHWPRYPELWMKLYELE LYCIILSLPPCLKISRRNQKQLTLFRLTLQNCHYQIIPPHILLDTGLIQLPVIWR SEQ ID NO: 38 >tr|A0A2Y9E587|A0A2Y9E587_TRIMA C->U-editing enzyme APOBEC-1 OS = Trichechus manatus latirostris OX = 127582 GN = LOC101361717 PE = 4 SV = 1 MTSEEADQRHSTMTSEKGPSTGDGTLRRRITPWEFEIFFDPRELRKETCLLYEIKWGTSHRI WRNSGQNTTKHAEVNFIEKFTSERNFCPSVSCSITWFLSWSPCWECSKAIREFLSQHPNVI LVIYVARLFHHMDQQNREGLRDLVLSGVTVQIMSVSEYGHCWRNFVNYPPGEEARWPRYP PLWMMLYALELHCIILGLPPCLKISRRRQSQLTLFSLTPQNCHYQMIPPHILLATGLIQPYVTW R SEQ ID NO: 39 >tr|G1LKL4|G1LKL4_AILME CMP/dCMP-type deaminase domain-containing protein OS = Ailuropoda melanoleuca OX = 9646 GN = APOBEC1 PE = 4 SV = 1 ISWSTGPSGGDATSRRRIEPWEFEVFFDPRQLRKEACLLYEIQWGTSRKIWRNSGKNTTNH VEINFIEKFTLERQYCPSIHCSVTWFLSWSPCWECSKAIRAFLSQHPSVTLVIYVARLFWHM EPQNRQGLRDLINSGVTIQIMSVPEYDHCWRNFVNYPPGKDHWPGYPVLWMKLYALELHC IILSLPPCLKISRRNQNQLTLFRLTLQNCHYQTIPPHVLLATGLIQLPVTWR SEQ ID NO: 40 >tr|A0A093PWR2|A0A093PWR2_9PASS C->U-editing enzyme APOBEC-1 (Fragment) OS = Manacus vitellinus OX = 328815 GN = N305_14278 PE = 4 SV = 1 RWKIQPKDFKRNYLPGQHPQWYLLYEIRWRNGSIWRNWFSNNRNQHAEVNFLENCFSDV PPAPCSITWFLSTSPCGKCSRRILEFLRTHRNVTLEIYAAKLFRHQDIRNRQGLCNLVMNGV TIHIMNLADYSYCWKRFVAY SEQ ID NO: 41 >sp|Q9EQP0|ABEC1_MESAU C->U-editing enzyme APOBEC-1 OS = Mesocricetus auratus OX = 10036 GN = APOBEC1 PE = 2 SV = 1 MSSETGPVVVDPTLRRRIEPHEFDAFFDQGELRKETCLLYEIRWGGRHNIWRHTGQNTSR HVEINFIEKFTSERYFYPSTRCSIVWFLSWSPCGECSKAITEFLSGHPNVTLFIYAARLYHHT DQRNRQGLRDLISRGVTIRIMTEQEYCYCWRNFVNYPPSNEVYWPRYPNLWMRLYALELY CIHLGLPPCLKIKRRHQYPLTFFRLNLQSCHYQRIPPHILWATGFI SEQ ID NO: 42 >tr|A0A2K6PRF3|A0A2K6PRF3_RHIRO CMP/dCMP-type deaminase domain-containing protein OS = Rhinopithecus roxellana OX = 61622 GN = APOBEC1 PE = 4 SV = 1 MSWKIWRSSGKNTTNHVEVNFIEKFTSERRFHSSISCSITWFLSWSPCWDCSQAIRKFLSQ HPGVTLVIYVARLFWHTDQQNRQGLRDLVNSGVTIQMMTASEYYHCWRNFVNYPPGEEA HWPRYPPLWMMLYALELHCIILSLPPCLKISRRWQNHLTFFRLRLQNCHYQTIPPHILLATGL IQPSVTWR SEQ ID NO: 43 >tr|A0A0D9RBS4|A0A0D9RBS4_CHLSB Apolipoprotein B mRNA editing enzyme catalytic subunit 1 OS = Chlorocebus sabaeus OX = 60711 GN = APOBEC1 PE = 4 SV = 1 MSRKIWRSSGKNTTNHVEVNFIEKLTSERRFHSSVSCSVTWFLSWSPCWECSQAIREFLS QHPGVTLVIYVARLFWHTDQQNRQGLRDLVNSGVTIQIMRASEYYHCWRNFVNYPPGEEA HWPRYPPLWMMLYALELHCIILSLPPCLKISRRWQNHLTFFRLHLQNCHYQTIPPHILLATGL IQPPVTWR SEQ ID NO: 44 >tr|A0A286XNR2|A0A286XNR2_CAVPO CMP/dCMP-type deaminase domain-containing protein OS = Cavia porcellus OX = 10141 GN = APOBEC1 PE = 4 SV = 1 MASGTGPSTGDATLRRRIEPWQFEAYFDPRQLRKEACMLSEVRWGASPRTWRESSLNTT SHVEINFIEKFTSGRSLRPAVRCSMTWFLSWSPCWECARAIREFLHQHPNVSLVIYVARLY WHVDEQNRQGLRDLVTSGVRVQIMSDSEYRHCWRNFVNFPPGQEAGWPRFPPMWTTLY ALELSCILLSLPPCLKISRRRQYRLIVFQLILQTCHYRAIPPQVLSAAELMHPLVAWC SEQ ID NO: 45 >tr|A0A2Y9HAT6|A0A2Y9HAT6_NEOSC C->U-editing enzyme APOBEC-1 OS = Neomonachus schauinslandi OX = 29088 GN = APOBEC1 PE = 4 SV = 1 MASDKGPSAGDATLRRRIKPWEFEVFFDPRELRKETCLLYEIQWGTSHKIWRNSGKNTAN HVEINFIEKFTSERQYCPSIRCSITWFLSWSPCWECSKAIRGFLSQHPSVTLVIYVARLFWH MDPQNRQGLRDLINSGVTIQIMRVPEYDHCWRNFVNYLPGKEDHWPRYPVLWMKLYALEL HCIILSLPPCLRISRRQNQLTLFTLTLQNCHYQMIPPHILLATGLIQVPVTWK SEQ ID NO: 46 >tr|A0A091XJL0|A0A091XJL0_OPIHO C->U-editing enzyme APOBEC-1 (Fragment) OS = Opisthocomus hoazin OX = 30419 GN = N306_09750 PE = 4 SV = 1 RWKVQPNDFKRNYLPGQHPKVVYILYEIRWSRGTIWRNWCTNNSTQHAEVNFLENCFKAM PSVSCSITWVLSTTPCGKCSKRIQDFLRIYPNVTLEIHAAKLFKHLDTRNREGLRNLAKDGVII HIMNLADYSYWWKRFVAY SEQ ID NO: 47 >tr|F6WR88|F6WR88_HORSE Apolipoprotein B mRNA editing enzyme catalytic subunit 1 OS = Equus caballus OX = 9796 GN = APOBEC1 PE = 4 SV = 2 MSHNIWRYSGKNTTKHVEINFIEKFTSERHLRPSISCSIVWFLSWSPCWECSKAIREFLS QHPNVTLVIYVARLFQHMDRLNRQGLRDLINSGVTIQIMRTSEYDHCWRNFVNYPPGKEAH WPRYPLLWMKLYALELHCIILSLPPCLMISRRCQNQLTFFRLTLQNCHYQMIPPHILLATGLV QLPVTWR SEQ ID NO: 48 >sp|P41238|ABEC1_HUMAN C->U-editing enzyme APOBEC-1 OS = Homo sapiens OX = 9606 GN = APOBEC1 PE = 1 SV = 3 MTSEKGPSTGDPTLRRRIEPWEFDVFYDPRELRKEACLLYEIKWGMSRKIWRSSGKNTTN HVEVNFIKKFTSERDFHPSMSCSITWFLSWSPCWECSQAIREFLSRHPGVTLVIYVARLFW HMDQQNRQGLRDLVNSGVTIQIMRASEYYHCWRNFVNYPPGDEAHWPQYPPLWMMLYA LELHCIILSLPPCLKISRRWQNHLTFFRLHLQNCHYQTIPPHILLATGLIHPSVAWR SEQ ID NO: 49 >tr|A0A091RU17|A0A091RU17_NESNO C->U-editing enzyme APOBEC-1 (Fragment) OS = Nestor notabilis OX = 176057 GN = N333_10787 PE = 4 SV = 1 RWKIQPNDFKRNYLPYQHPKVVCLLYEIRWNRGTIWRSWCSNNSTQHAEVNFLENCFKAK PSVSCSITWVLSTTPCGECSRRILDFLSVYPNVTLKIYAAKLFKHLDNRNRQGLWNLANNRV IIRIMNLEDYNYYWKRFVAY SEQ ID NO: 50 >tr|A0A091IWL9|A0A091IWL9_EGRGA C->U-editing enzyme APOBEC-1 (Fragment) OS = Egretta garzetta OX = 188379 GN = Z169_O8812 PE = 4 SV = 1 RWKIQPNDFKRNYLPGQHPKVVYLLYEIRWSRGTIWRNWCSNNSTQHAEVNFLENCFKAM PSVSCSITWVLSTTPCGKCSRRILEFLRVHPSVTLEIYAAKLFKHLDIRNRQGLRNLAMNGVII HIMNLADYSYWWKIFVAY SEQ ID NO: 51 >tr|A0A2K5DG70|A0A2K5DG70_AOTNA CMP/dCMP-type deaminase domain-containing protein OS = Aotus nancymaae OX = 37293 GN = APOBEC1 PE = 4 SV = 1 MTPEEEVQRQSTMTSERGPSTGDPTLRRRIEPWEFCISYDPKELCKETCLLYEIKWGTSWK IWRSSGKNTTNHVEVNFIEKFMSERHFHSSISCSITWFLSWSPCWECSQAIREFLSRHPGV TLVIYVARLFQHMDRQNRQGLRDLVNSGVTIQIMRASEYYHCWRNFVNYPPGEEAHWPRY PPLWMMLYALELHCIILGLPPCLKISRRWQNRLTFFRLHLQNCHYQMIPQHILFATGLIQPPV TWR SEQ ID NO: 52 >sp|P51908|ABEC1_MOUSE C->U-editing enzyme APOBEC-1 OS = Mus I OX = 10090GN = Apobec1 PE = 1 SV = 1 MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSVWRHTSQNTSN HVEVNFLEKFTTERYFRPNTRCSITWFLSWSPCGECSRAITEFLSRHPYVTLFIYIARLYHHT DQRNRQGLRDLISSGVTIQIMTEQEYCYCWRNFVNYPPSNEAYWPRYPHLWVKLYVLELY CIILGLPPCLKILRRKQPQLTFFTITLQTCHYQRIPPHLLWATGLK SEQ ID NO: 53 >tr|G5BPM8|G5BPM8_HETGA C->U-editing enzyme APOBEC-1 (Fragment) OS = Heterocephalus glaber OX = 10181 GN = GW7_17308 PE = 4 SV = 1 RRRIEPWQFEASFDPRQLRRETCLLSEVRWGTSPRAWRGCSLNTARHAEVSFMDRLTSE GRLRGPVRCSITWFLSWSPCGACAQAIGEFLRQHPNVSLVIYIARLFWHVDEQNRQGLRDL VTRGVRMQVMSDPEFAHCWRNFVNYSPGQEARWPQVPPWVTWLYSLELHCILLNLPPCL KISRRHHNQLTFFQLILQNCHYQAIPSPVLLASGLIHPFVTW SEQ ID NO: 54 >tr|A0A091QEK6|A0A091QEK6_MERNU C->U-editing enzyme APOBEC-1 (Fragment) OS = Merops nubicus OX = 57421 GN = N331_01832 PE = 4 SV = 1 RWKIEPDEFKTNYSPDHRPRVVYLLYEIRWRRGTIWRNWCSNNIDQHAEVNFLENCFKAK PSVSCSITWFLSTAPCAKCSRRILKFLTAHPKVTLEIYAAKLFRHLEIRNRQGLMDLAVN GVILRIMNLADYSYCWKQFVAY SEQ ID NO: 55 >tr|A0A093LP85|A0A093LP85_FULGA C->U-editing enzyme APOBEC-1 (Fragment) OS = Fulmarus glacialis OX = 30455 GN = N327_13724 PE = 4 SV = 1 RWKIQPNDFKRNFLPSKYPKVVYLLYEIRWSSGTIWRSWCSNNSTQHAEVNFLENCFKAM PSVSCSITWVLPITPCGKCSKKILEFLSVHPNVTLEIYAAKLFRHLDIRNQQGLRNLAMN GVIIRIMNLADYSYSWKRFVAY SEQ ID NO: 56 >tr|G1QZV0|G1QZV0_NOMLE CMP/dCMP-type deaminase domain-containing protein OS = Nomascus leucogenys OX = 61853 GN = APOBEC1 PE = 4 SV = 1 MTSEKGPSTGDPTLRRRIEPWEFDVFYDPRELRKEACLLYEIKWGMSQKIWRSSGKNTTN HVEVNFIKKFTSEGRFQSSISCSITWFLSWSPCWECSQAIREFLSQHPGVTLVIYVARLF WHMDQQNRQGLRDLVNSGVTIQIMRASEYYHCWRNFVNYPPGDEAHWPRYPPLWMMLY AL ELHCIILSLPPCLKISRRWQNHLTFFRLHLQNCHYQTIPPHILLATGLIHPSVTWR SEQ ID NO: 57 >tr|A0A096MWB4|A0A096MWB4_PAPAN CMP/dCMP-type deaminase domain-containing protein OS = Papio anubis OX = 9555 GN = APOBEC1 PE = 4 SV = 2 MTSEKGPSTGDPTLRRRIEPWEFDIFYDPRELRKEACLLYEIKWGMSPKIWRSSGKNTTN HVEVNFIEKLTSERRFHSSISCSITWFLSWSPCWECSQAIREFLSQHPGVTLVIYVARLF WHTDQQNRQGLRDLVNSGVTIQIMTASEYYHCWRNFVNYPPGEEAHWPRYPPLWMMLY AL ELHCIILSLPPCLKISRRWQNHLTFFRLHLQNCHYQTIPPHILLATGLIQPSVTWR SEQ ID NO: 58 >sp|Q9TUI7|ABEC1_MONDO C->U-editing enzyme APOBEC-1 OS = Monodelphis domestica OX = 13616 GN = APOBEC1 PE = 1 SV = 1 MNSKTGPSVGDATLRRRIKPWEFVAFFNPQELRKETCLLYEIKWGNQNIWRHSNQNTSQH AEINFMEKFTAERHFNSSVRCSITWFLSWSPCWECSKAIRKFLDHYPNVTLAIFISRLYW HMDQQHRQGLKELVHSGVTIQIMSYSEYHYCWRNFVDYPQGEEDYWPKYPYLWIMLYVLE LHCIILGLPPCLKISGSHSNQLALFSLDLQDCHYQKIPYNVLVATGLVQPFVTWR SEQ ID NO: 59 >tr|A0A1S3FTE2|A0A1S3FTE2_DIPOR C->U-editing enzyme APOBEC-1 OS = Dipodomys ordii OX = 10020 GN = Apobec1 PE = 4 SV = 1 MHHSARLPPNCIVSRYANAPWTVLPLPLPPTEAPATGDDTLRRRIEPWEFEAFFNPQELR REACLLYQITWSSHKVWRETAKNTVDSHVEVNFIQNLTAGRYCRPSTRCSILWFLSWSPC SSCSKAIRLFLSQHPGVSLVIYVARLFQHMDPQNRQGLRELIHSGVTIQVMRPQEYDYCWK NFVNYPPGQEEHWPRYPVQCMTLYNLELYCIIHNLPPCVRISKQRQSQLAFFSLGLENVHY QRIPPPLLLLTGLVFVFPWK SEQ ID NO: 60 >tr|A0A2U3WPA5|AO0A2U3WPA5_ODORO C->U-editing enzyme APOBEC-1 OS = Odobenus rosmarus divergens OX = 9708 GN = APOBEC1 PE = 4 SV = 1 MASDKGPSAGDATLRRRIEPWEFEVFFDPRELRKEACLLYEIQWGTSHKIWRNSGKNTSN H VEIN FI EKFTSERQYCPSIHCSITWFLSWSPCWECSEAIRGFLSQHPSVTLVIYVARLFWH MDPQNRQGLRDLINSGVTIQIMRVPEYDHCWRNFVNYPPGKEDHWPRYPVLWMKLYALEL HCIILSLPPCLRISRRQNQLTLFRLTLQNCHYQMIPPHILLATGLIQVPVTWK SEQ ID NO: 61 >tr|A0A1V4JAP2|A0A1V4JAP2_PATFA C->U-editing enzyme APOBEC-1 OS = Patagioenas fasciata monilis OX = 372326 GN = APOBEC1 PE = 4 SV = 1 MRRKKPSGMYISKRALKDNFDPHKFPHDTYLLCKLQWGDTGRSWIHWIRKDRYHAEVYFL EKIFKMRRSKNYVNCSITWYLSWSPCVRCCCEILNFLEKHSYVNIDIYVARLYKIQNSEVREG LKKLVSSKKVTIAVMEIKDYTYCWKNFIQGDADDDSWTVDFQSAITKNRLKLKDVFEFLKSH PNVTLEIYAAKLFKHLDIRNREGLRNLAKNGVIIHIMNLADYSYWWKIFVTRQHGEDDYLPWS FALHIFLNCIEFQQILLVSRHLKESLRVKSNEKAQEKEVWRIPAMVLAEMIVGKMNRDLMLHE QRANRARNCKGLWCYIVPL SEQ ID NO: 62 >tr|A0A2K5JKV4|A0A2K5JKV4_COLAP CMP/dCMP-type deaminase domain-containing protein OS = Colobus angolensis palliatus OX = 336983 GN = APOBEC1 PE = 4 SV = 1 PSTGDPTLRRRIEPWEFDIFYDPRELRKEACLLYEIKWGMSQKIWRSSGKNTTNHVEVNFIE KLTSERRFHSSVSCSITWFLSWSPCWECSQAIREFLSQHPGVTLVIYVARLFWHTDQQNRQ GLRDLVNSGVTIQMMTASEYYHCWRNFVNYPPGEEAHWPRYPPLWMMLYALELHCIILSL PPCLKISRRWQNHLTFFRLHLQNCHYQTIPPHILLATGLIQPSVTWR SEQ ID NO: 63 >tr|A0A1U7U8J6|A0A1U7U8J6_TARSY LOW QUALITY PROTEIN: C->U-editing enzyme APOBEC-1 OS = Tarsius syrichta OX = 1868482 GN = APOBEC1 PE = 4 SV = 2 MLTALMEEVQDTMRFGRRAFFLSNSVGIWVLFDISISXSTGPSMGDPTLRRRIEPWEFEVLF DPRELRKEACLLYEIKWGTSCKIWRNSGKNTSNHAEVNFLEKFTSERHFCSSTSYSITWFLS WSPCWECSRAIREFLSQHPRVTLVIYVARLFWHMEPQNRQGLRDLINSGVTIQIMRDSGKS NKQIIRIVCERTW SEQ ID NO: 64 >tr|F1SLW4|F1SLW4_PIG CMP/dCMP-type deaminase domain-containing protein OS = Sus scrota OX = 9823 GN = APOBEC1 PE = 4 SV = 2 MASDRGPSAGDATSRRRIEPWEFEVFFDPRELRKETCLLYELQWGRSRDTWRHTGKNTT NHVERNFLAKITSERHFHPSVHCSIVWFLSWSPCWECSEAIREFLDQHPSVTLVIYVARLFQ HMDPQNRQGLRDLVNHGVTIQIMGAPEYDYCWRNFVNYPPGKEAHWPRFPPVWMTLYAL ELHCIILGLPPCLKISRRCQNQLTFFRLTLQNCHYQTIPPHILLATGLIQLPVIYR SEQ ID NO: 65 >tr|A0A2K6BGI5|A0A2K6BGI5_MACNE CMP/dCMP-type deaminase domain-containing protein OS = Macaca nemestrina OX = 9545 GN = APOBEC1 PE = 4 SV = 1 MTSEKGPSTGDPTLRRRIEPWEFDIFYDPRELRKEACLLYEIKWGMSPKIWRSSGKNTTNH VEVNFIEKLTSERRFHSSISCSITWFLSWSPCWECSQAIREFLSQHPGVTLVIYVARLFWHT DQQNRQGLRDLVNSGVTIQIMRASEYYHCWRNFVNYLPGEEAHWPRYPPLWMMLYALEL HCIILSLPPCLKISRRWQNHLTFFRLHLQNCHYQMIPPHILLATGLIQPSVTWR SEQ ID NO: 66 >sp|P47855|ABEC1_RABIT C->U-editing enzyme APOBEC-1 OS = Oryctolagus cuniculus OX = 9986 GN = APOBEC1 PE = 1 SV = 1 MASEKGPSNKDYTLRRRIEPWEFEVFFDPQELRKEACLLYEIKWGASSKTWRSSGKNTTN HVEVNFLEKLTSEGRLGPSTCCSITWFLSWSPCWECSMAIREFLSQHPGVTLIIFVARLFQH MDRRNRQGLKDLVTSGVTVRVMSVSEYCYCWENFVNYPPGKAAQWPRYPPRWMLMYAL ELYCIILGLPPCLKISRRHQKQLTFFSLTPQYCHYKMIPPYILLATGLLQPSVPWR SEQ ID NO: 67 >sp|P38483|ABEC1_RAT C->U-editing enzyme APOBEC-1 OS = Rattus norvegicus OX = 10116GN = Apobec1 PE = 1 SV = 1 MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKH VEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHAD PRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCI ILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLK SEQ ID NO: 68 >tr|A0A091M4D7|A0A091M4D7_CARIC C->U-editing enzyme APOBEC-1 (Fragment) OS = Cariama cristata OX = 54380 GN = N322_12137 PE = 4 SV = 1 RWKIQPDDFKRNYLPGQHPEVVYLLYEIKWNSGTIWRNWCSNNPTQHAEVNFLENHFNVM SSVSCSITWGISTTPCGKCSRRILEFLTTHPNVTLEIYAAKLFKHLDIRNRQGLRNLAMNGVVI CIMNLADYSYFWKTFVAY SEQ ID NO: 69 >tr|A0A093F3R4|A0A093F3R4_GAVST C->U-editing enzyme APOBEC-1 (Fragment) OS = Gavia stellata OX = 37040 GN = N328_12441 PE = 4 SV = 1 RWKIQPNDFKRNYLPAQHPKVVYLLYEIRWSRGTIWRNWCSNNSTQHAEVNFLENCFKAM PSVSCSITWFLSTTPCGKCSRRILTFLREHPNVTLEIYAAKLFKHLDVRNQQGLRNLDRNGVI IRIMNFADYSYCWKRFVAY SEQ ID NO: 70 >tr|G7N5W0|G7N5W0_MACMU C->U-editing enzyme APOBEC-1 (Fragment) OS = Macaca mulatta OX = 9544 GN = EGK_03318 PE = 4 SV = 1 GPSTGDPTLRRRIEPWEFDIFYDPRELRKEACLLYEIKWGMSPKIWRSSGKNTTNHVEVNFI EKLTSERRFHSSISCSITWFLSWSPCWECSQAIREFLSQHPGVTLVIYVARLFWHTDQQNR QGLRDLVNSGVTIQIMRASEYYHCWRNFVNYPPGEEAHWPRYPPLWMMLYALELHCIILSL PPCLKISRRWQNHLTFFRLHLQNCHYQMIPPHILLATGLIQPSVTWR SEQ ID NO: 71 >tr|A0A091MEP8|A0A091MEP8_9PASS C->U-editing enzyme APOBEC-1 (Fragment) OS = Acanthisitta chloris OX = 57068 GN = N310_12928 PE = 4 SV = 1 RWKIQPNDFQRNYLPDQHPQAVYLLYEFRWRRGSIWRKWCSNNRAQHAEVNFLENCFNG IPPVPCSITWFLSTTPCGNCSRRILEFLRLHPNVTLEIYAAKLFRHTDIRNRKGLYNLAMNGVII RIMNLADYSYCWRNFVAY SEQ ID NO: 72 >tr|A0A2I0LXZ8|A0A2I0LXZ8_COLLI Apolipoprotein B mRNA editing enzyme, catalytic polypeptide 1 OS = Columba livia OX = 8932 GN = APOBEC1 PE = 4 SV = 1 MAAVTNRDSACRENNQRWKIQPNDFRRNYLPDKQPRVVYLLYEIRWRRGTIWRNWCSNN PNQHAEVNFLKNYFNAMPSVSCSITWVLSTTPCGKCSIKIMEFLKLHPNVTLEIYAAKLFKHL DIRNREGLRNLAKNGVIIHIMNLADYSYWWKIFVTRQHGEEDYLPWSFALHIFLNCIEFQQILL GLPPLLPNFKY SEQ ID NO: 73 >tr|W5NVH9|W5NVH9_SHEEP CMP/dCMP-type deaminase domain-containing protein OS = Ovis aries OX = 9940 GN = APOBEC1 PE = 4 SV = 1 MASDRGPPAGDPTLRRRIEPLEFEFSFDPRNFCKEAYLLYEIQWGNSRDVWRHSGKNTTK HVERNFIEKIASERHFRPSISCSISWYLSWSPCWECSKAIREFLNQHPNVTLVIYIARLFQHM DPQNRQGLKDLFHSGVTIQVMRDPEYDYCWRNFVNYPQGKEAHWPRYPPLWMNLYALEL YCIISGLPPCLQISRRHQNQLRVFRLIPQNCHYQMIPPCILLATGMIQLPVTWRWIE SEQ ID NO: 74 >tr|H0XVG8|H0XVG8_OTOGA CMP/dCMP-type deaminase domain-containing protein OS = Otolemur garnettii OX = 30611 GN = APOBEC1 PE = 4 SV = 1 ISWSTGISTGDPTLRRRIEPWEFEVFFDPRELRKETCLLYEIKWGTSHKIWRSTARNTTS HAEMNFIEKFTSERCSDAPISCSITWFLSWSPCWECSKAIREFVSRHPSVTLVIYVARLY WHMDQQNRQGLRDLISSGVTVQIMRVSEYCHCWRNFVNYLPGKEAHCPRCPPLWMTLYA LELHCIILSLPPCLKISRGHQNQLTLFRLTLQNCHYQTIPPHVLLATGLIQPYVTWR SEQ ID NO: 75 >tr|A0A2B4RXQ3|A0A2B4RXQ3_STYPI C->U-editing enzyme APOBEC-1 OS = Stylophora pistillata OX = 50429 GN = APOBEC1 PE = 4 SV = 1 MASVTELRTPDDFLAELLWTGVTGRTWPNRTFLIVSIKAKDGKPIFGKRFKNRYPEHAEI IMLRNSNFSDVVEKNHDIDITLTLNYSPCSSCACILKEFYVNNSNIKCFTIQFSFIYYKE DMKNKTGLQNLEEAGVTLQAMNAESWREVGIDLESFTPEDKEKINKRDKDTANDLNEVLSS KQDQDASVDELSSQLNAKLRAKET SEQ ID NO: 76 >tr|A0A2K5L2J6|A0A2K5L2J6_CERAT CMP/dCMP-type deaminase domain-containing protein OS = Cercocebus atys OX = 9531 GN = APOBEC1 PE = 4 SV = 1 MTPEEEVQRQSTMTSEKGPSTGDPTLRRRIEPWEFDIFYDPRELRKEACLLYEIKWGMSPK IWRSSGKNTTNHVEVNFIEKLTSERRFHSSISCSITWFLSWSPCWECSQAIREFLSQHPGVT LVIYVARLFWHTDQQNRQGLRDLVNSGVTIQIMRASEYYHCWRNFVNYPPGEEAHWPRYP PLWMMLYALELHCIILSLPPCLKISRRQQNHLTFFRLHLQNCHYQTIPPHILLATGLIQPSVTW R SEQ ID NO: 77 >tr|A0A2Y9T649|A0A2Y9T649_PHYMC C->U-editing enzyme APOBEC-1 isoform X1 OS = Physeter macrocephalus OX = 9755 GN = APOBEC1 PE = 4 SV = 1 MIICWSTGPSAGDATSRRRIEPWEFEVSFDPREFCKEARLLYEIKWGKSQDVWRHSGKNT TKHVECNFIEKMTSERHFHPSISCCIIWFLSWSPCWECSKAIREFLNQHPSVTLVIYIARLFQ HTDPQNRQGLRDLIHSGVTLQIMGPPEYDYCWRNFVNYPPGKEAHWPRYPPLWMKLYAL ELHCIILGLPPCLKISRRCQNQLTWFRLILQNCHYQMIPPHILLGTGLIQLPVAWR SEQ ID NO: 78 >tr|H2NGDO|H2NGDO_PONAB Apolipoprotein B mRNA editing enzyme catalytic subunit 1 OS = Pongo abelii OX = 9601 GN = APOBEC1 PE = 4 SV = 1 MTPEEEVQRQSTMTSEKGPSTGDPTLRRRIESWEFDVFYDPRELRKETCLLYEIKWGMSR KIWRSSGKNTTNHVEVNFIKKFTSERRFHSSISCSITWFLSWSPCWECSQAIREFLSQHPGV TLVIYVARLFWHMDQRNRQGLRDLVNSGVTIQIMRASEYYHCWRNFVNYPPGDEAHWPQ YPPLWMMLYALELHCIILSLPPCLKISRRWQNHLAFFRLHLQNCHYQTIPPHILLATGLIHPSV TWK SEQ ID NO: 79 >tr|A0A093JI54|A0A093JI54_EURHL C->U-editing enzyme APOBEC-1 (Fragment) OS = Eurypyga helias OX = 54383 GN = N326_10046 PE = 4 SV = 1 RWKIQPNDFKRNYMPSQYPKVVYLLYEIRWSRGTVWRNWCSNSFTQHAEVNFLENYFKP MPSVSCSITWVLSTTPCGKCSRRILEFLRVHPNVTLEIYAAKLFKHLDIRNRQGLRDLAMNG VTIRIMNLADYSFCWKRFVAY SEQ ID NO: 80 >tr|G3W4H|G3W4H_SARHA Apolipoprotein B mRNA editing enzyme catalytic subunit 1 OS = Sarcophilus harrisii OX = 9305 GN = APOBEC1 PE = 4 SV = 1 MGDATLRRRIKSWEFEAFFNPQELRKETCLLYEIKWGASHNIWRSSNQNTTQHAEINFMEK FTSERNFKPSVKCSITWFLSWSPCWRCSKAIREFLNQYPNVTLVIFVSRLYWHMEQQHRQ ELKELVCSGVTIQIMNYSEYRHCWRNFVDYLPEEEDHWPKYPTLWIMLYVLELHCIILGLPP CLKISVRHSDQLVLFSLDLQDCHYQKIPYHVLVATGIIRPFVTWR SEQ ID NO: 81 >tr|A0A2U3Y3M5|A0A2U3Y3M5_LEPWE c->U-editing enzyme APOBEC-1 OS = Leptonychotes weddellii OX = 9713 GN = APOBEC1 PE = 4 SV = 1 MASDKGPSAGDATLRRRIKPWEFEVFFDPRELRKETCLLYEIQWGTSHKIWRNSGKNTAN HVEINFIEKFTSERQYCPSIRCSITWFLSWSPCWECSKAIRGFLSQHPSVTLVIYVARLFWH MDPQNRQGLRDLINSGVTIQIMRVPEYDHCWRNFVNYLPGKEDHWPRYPVLWMKLYALEL HCIILPIEMPGKIRDAPNNMEIFSLFVGRYIPKTKFHVTCLLSDVRNDDSHLDKTAPKWIRFDS LQPVASDPSAEHWKMKLPGRDDKTAVVVGTVTEDVACAQGAKLYLCALRVHGHAQRHFL KGRDEILALDQLALDSPQGLWRQPDLRSHPLKG SEQ ID NO: 82 >tr|A0A1S3AN78|A0A1S3AN78_ERIEU C->U-editing enzyme APOBEC-1-like OS = Erinaceus europaeus OX = 9365 GN = LOC103126721 PE = 4 SV = 1 RRIEPWEFEDFFDPRQFRPETCLLYEVRWGSSRNAWRSTARNTTRHAEVNFLERFAAERH FDKPVSCSITWFLSWSPCWECSQAIGAFLSQHPQVTLAIHVTRLFHHEDEQNRQGLRDLLA RGVTLQVMGDSEYAHCWRTFVNSPPGAEGHYPRYPSDFTRLYALELHCIILGLPPCLEILRR YQNQFTLFRLVPQNCHYQMIPHLNFFVVRHYFF SEQ ID NO: 83 >tr|A0A091PSV3|A0A091PSV3_HALAL C->U-editing enzyme APOBEC-1 (Fragment) OS = Haliaeetus albicilla OX = 8969 GN = N329_07103 PE = 4 SV = 1 RWKLQPNDFKRNYLPGQHPKVVYLLYEIRWSRGTIWRNWCSNNSTQHAEVNFLENCFKAT PSVSCSITWVLSTTPCGKCSRRILEFLRVHPNVTLEIYAAKLFKHLDIRNRKGLRDLAMNGVII RIMNLSDYSYCWKTFVAY SEQ ID NO: 84 >tr|F7F6M6|F7F6M6_CALJA Apolipoprotein B mRNA editing enzyme catalytic subunit 1 OS = Callithrix jacchus OX = 9483 GN = APOBEC1 PE = 4 SV = 2 RRIEPWEFYISYDPKELCKETCLLYEIKWGMSWKIWRSSGKNTTNHVEINFIEKFTSERH FHLSVSCSITWFLSWSPCWECSQAIREFLSQHPGVTLVIYVARLFQHMDQQNRQGLRDLV NSGVTIQMMTVSEYYHCWRNFVNYPPGEEAHWPRHPPLWLMLYALELHCIILGLPPCLKIS RRRQNRLTFFRLHLQNCHYQMIPRHILLATGLIQPSVTWR SEQ ID NO: 85 >tr|L8IDZ0|L8IDZ0_9CETA C->U-editing enzyme APOBEC-1 OS = Bos mutus OX = 72004 GN = M91_02456 PE = 4 SV = 1 MIISWSTGPPAGDPTLRRRIEPWEFEFSFDPRKFCKEACLLYEIQWGNNRDVWRHSGKNTT KHVERNFIEKIASERYFCPSIRCFIFWYLSWSPCWECSKAIREFLNQHPNVTLVIYIARLFQH MDPQNRQGLKDLVQSGVTIQVMRAPEYEYCWRNFVNYPRGKEAHWPRYPPLWMNLYAL ELYCIILGLPPCLHISRRYQNQLIVFRLTLQNCHYQMIPPYILLATGMVQIPMTWR SEQ ID NO: 86 >tr|A0A093CIQ8|A0A093CIQ8_9AVES C->U-editing enzyme APOBEC-1 (Fragment) OS = Pterocles gutturalis OX = 240206 GN = N339_03265 PE = 4 SV = 1 RWKIQPNYFKINNLPGQHPRVVCLLYAIRWSRSTLWKSWCSNNSTQHAEVNFLENCFKGN PSVFCFMTWPFFHTTPHGKCCRRTPEFLGVHPNVTLKIRAAKLFKHLDRYNQQGLRNVAM NGVVIRIINL SEQ ID NO: 87 >sp|Q9GZX7|AICDA_HUMAN Single-stranded DNA cytosine deaminase OS = Homo sapiens OX = 9606 GN = AICDA PE = 1 SV = 1 MDSLLMNRRKFLYQFKNVRWAKGRRETYLCYVVKRRDSATSFSLDFGYLRNKNGCHVELL FLRYISDWDLDPGRCYRVTWFTSWSPCYDCARHVADFLRGNPNLSLRIFTARLYFCEDRKA EPEGLRRLHRAGVQIAIMTFKDYFYCWNTFVENHERTFKAWEGLHENSVRLSRQLRRILLP LYEVDDLRDAFRTLGL SEQ ID NO: 88 >sp|Q9Y235|ABEC2_HUMAN C->U-editing enzyme APOBEC-2 OS = Homo sapiens OX = 9606 GN = APOBEC2 PE = 1 SV = 1 MAQKEEAAVATEAASQNGEDLENLDDPEKLKELIELPPFEIVTGERLPANFFKFQFRNVEYS SGRNKTFLCYVVEAQGKGGQVQASRGYLEDEHAAAHAEEAFFNTILPAFDPALRYNVTWY VSSSPCAACADRIIKTLSKTKNLRLLILVGRLFMWEEPEIQAALKKLKEAGCKLRIMKPQDFE YVWQNFVEQEEGESKAFQPWEDIQENFLYYEEKLADILK SEQ ID NO: 89 >sp|P31941|ABC3A_HUMAN DNA dC->dll-editing enzyme APOBEC-3A OS = Homo sapiens OX = 9606 GN = APOBEC3A PE = 1 SV = 3 MEASPASGPRHLMDPHIFTSNFNNGIGRHKTYLCYEVERLDNGTSVKMDQHRGFLHNQAK NLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISWSPCFSWGCAGEVRAFLQENTHVR LRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWDTFVDHQGCPFQPWDGLDE HSQALSGRLRAILQNQGN SEQ ID NO: 90 >sp|Q9UH17|ABC3B_HUMAN DNA dC->dU-editing enzyme APOBEC-3B OS = Homo sapiens OX = 9606 GN = APOBEC3B PE = 1 SV = 1 MNPQIRNPMERMYRDTFYDNFENEPILYGRSYTWLCYEVKIKRGRSNLLWDTGVFRGQVY FKPQYHAEMCFLSWFCGNQLPAYKCFQITWFVSWTPCPDCVAKLAEFLSEHPNVTLTISAA RLYYYWERDYRRALCRLSQAGARVTIMDYEEFAYCWENFVYNEGQQFMPWYKFDENYAF LHRTLKEILRYLMDPDTFTFNFNNDPLVLRRRQTYLCYEVERLDNGTWVLMDQHMGFLCNE AKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISWSPCFSWGCAGEVRAFLQENTH VRLRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFEYCWDTFVYRQGCPFQPWDGLE EHSQALSGRLRAILQNQGN SEQ ID NO: 91 >sp|Q9NRW3|ABC3C_HUMAN DNA dC->dU-editing enzyme APOBEC-3C OS = Homo sapiens OX = 9606 GN = APOBEC3C PE = 1 SV = 2 MNPQIRNPMKAMYPGTFYFQFKNLWEANDRNETWLCFTVEGIKRRSVVSWKTGVFRNQV DSETHCHAERCFLSWFCDDILSPNTKYQVTWYTSWSPCPDCAGEVAEFLARHSNVNLTIFT ARLYYFQYPCYQEGLRSLSQEGVAVEIMDYEDFKYCWENFVYNDNEPFKPWKGLKTNFRL LKRRLRESLQ SEQ ID NO: 92 >sp|Q96AK3|ABC3D_HUMAN DNA dC->dU-editing enzyme APOBEC-3D OS = Homo sapiens OX = 9606 GN = APOBEC3D PE = 1 SV = 1 MNPQIRNPMERMYRDTFYDNFENEPILYGRSYTWLCYEVKIKRGRSNLLWDTGVFRGPVL PKRQSNHRQEVYFRFENHAEMCFLSWFCGNRLPANRRFQITWFVSWNPCLPCVVKVTKF LAEHPNVTLTISAARLYYYRDRDWRWVLLRLHKAGARVKIMDYEDFAYCWENFVCNEGQP FMPWYKFDDNYASLHRTLKEILRNPMEAMYPHIFYFHFKNLLKACGRNESWLCFTMEVTKH HSAVFRKRGVFRNQVDPETHCHAERCFLSWFCDDILSPNTNYEVTWYTSWSPCPECAGE VAEFLARHSNVNLTIFTARLCYFWDTDYQEGLCSLSQEGASVKIMGYKDFVSCWKNFVYSD DEPFKPWKGLQTNFRLLKRRLREILQ SEQ ID NO: 93 >sp|Q8IUX4|ABC3F_HUMAN DNA dC->dU-editing enzyme APOBEC-3F OS = Homo sapiens OX = 9606 GN = APOBEC3F PE = 1 SV = 3 MKPHFRNTVERMYRDTFSYNFYNRPILSRRNTVWLCYEVKTKGPSRPRLDAKIFRGQVYS QPEHHAEMCFLSWFCGNQLPAYKCFQITWFVSWTPCPDCVAKLAEFLAEHPNVTLTISAAR LYYYWERDYRRALCRLSQAGARVKIMDDEEFAYCWENFVYSEGQPFMPWYKFDDNYAFL HRTLKEILRNPMEAMYPHIFYFHFKNLRKAYGRNESWLCFTMEWKHHSPVSWKRGVFRN QVDPETHCHAERCFLSWFCDDILSPNTNYEVTWYTSWSPCPECAGEVAEFLARHSNVNLTI FTARLYYFWDTDYQEGLRSLSQEGASVEIMGYKDFKYCWENFVYNDDEPFKPWKGLKYNF LFLDSKLQEILE SEQ ID NO: 94 >sp|Q9HC16|ABC3G_HUMAN DNA dC->dU-editing enzyme APOBEC-3G OS = Homo sapiens OX = 9606 GN = APOBEC3G PE = 1 SV = 1 MKPHFRNTVERMYRDTFSYNFYNRPILSRRNTVWLCYEVKTKGPSRPPLDAKIFRGQVYSE LKYHPEMRFFHWFSKWRKLHRDQEYEVTWYISWSPCTKCTRDMATFLAEDPKVTLTIFVA RLYYFWDPDYQEALRSLCQKRDGPRATMKIMNYDEFQHCWSKFVYSQRELFEPWNNLPK YYILLHIMLGEILRHSMDPPTFTFNFNNEPWVRGRHETYLCYEVERMHNDTWVLLNQRRGF LCNQAPHKHGFLEGRHAELCFLDVIPFWKLDLDQDYRVTCFTSWSPCFSCAQEMAKFISKN KHVSLCIFTARIYDDQGRCQEGLRTLAEAGAKISIMTYSEFKHCWDTFVDHQGCPFQPWDG LDEHSQDLSGRLRAILQNQEN SEQ ID NO: 95 >sp|Q6NTF7|ABC3H_HUMAN DNA dC->dU-editing enzyme APOBEC-3H OS = Homo sapiens OX = 9606 GN = APOBEC3H PE = 1 SV = 4 MALLTAETFRLQFNNKRRLRRPYYPRKALLCYQLTPQNGSTPTRGYFENKKKCHAEICFINE IKSMGLDETQCYQVTCYLTWSPCSSCAWELVDFIKAHDHLNLGIFASRLYYHWCKPQQKGL RLLCGSQVPVEVMGFPEFADCWENFVDHEKPLSFNPYKMLEELDKNSRAIKRRLERIKIPG VRAQGRYMDILCDAEV Petromyzon marinus cytosine deaminase (pmCDA1), Genbank ABO15149.1 SEQ ID NO: 96 MTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGERRACFWGYAVNKPQSGT ERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKILEWYNQELRGNGHTLKIWA CKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIFIQSSHNQLNENRWLEKTLKR AEKRRSELSIMIQVKILHTTKSPAV Petromyzon marinus cytosine deaminase (pmCDA1) R187W, as used in Target-AID, SEQ ID NO: 97 MTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGERRACFWGYAVNKPQSGT ERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKILEWYNQELRGNGHTLKIWA CKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIFIQSSHNQLNENRWLEKTLKR AEKWRSELSIMIQVKILHTTKSPAV E. coli TadA, SEQ ID NO: 98 MSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGWNRPIGRHDPTAHAEI MALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSRIGRVVFGARDAKTGAAGSLMDVL HHPGMNHRVEITEGILADECAALLSDFFRMRRQEIKAQKKAQSSTD S. aureus TadA, SEQ ID NO: 99 MTNDIYFMTLAIEEAKKAAQLGEVPIGAIITKDDEVIARAHNLRETLQQPTAHAEHIAIERAAKV LGSWRLEGCTLYVTLEPCVMCAGTIVMSRIPRVVYGADDPKGGCSGSLMNLLQQSNFNHR AIVDKGVLKEACSTLLTTFFKNLRANKKSTN S. pyogenes TadA, SEQ ID NO: 100 MPYSLEEQTYFMQEALKEAEKSLQKAEIPIGCVIVKDGEIIGRGHNAREESNQAIMHAEMMAI NEANAHEGNWRLLDTTLFVTIEPCVMCSGAIGLARIPHVIYGASNQKFGGADSLYQILTDER LNHRVQVERGLLAADCANIMQTFFRQGRERKKIAKHLIKEQSDPFD S. typhi TadA, SEQ ID NO: 101 MSDVELDHEYWMRHALTLAKRAWDEREVPVGAVLVHNHRVIGEGWNRPIGRHDPTAHAEI MALRQGGLVLQNYRLLDTTLYVTLEPCVMCAGAMVHSRIGRWFGARDAKTGAAGSLIDVL HHPGMNHRVEIIEGVLRDECATLLSDFFRMRRQEIKALKKADRAEGAGPAV A. aeolicus TadA, SEQ ID NO: 102 MGKEYFLKVALREAKRAFEKGEVPVGAIIVKEGEIISKAHNSVEELKDPTAHAEMLAIKEACR RLNTKYLEGCELYVTLEPCIMCSYALVLSRIEKVIFSALDKKHGGVVSVFNILDEPTLNHRVK WEYYPLEEASELLSEFFKKLRNNII S. pombe TAD2, SEQ ID NO: 103 MAGDSVKSAIIGIAGGPFSGKTQLCEQLLERLKSSAPSTFSKLIHLTSFLYPNSVDRYALSSY DIEAFKKVLSLISQGAEKICLPDGSCIKLPVDQNRIILIEGYYLLLPELLPYYTSKIFVYEDADTR LERCVLQRVKAEKGDLTKVLNDFVTLSKPAYDSSIHPTRENADIILPQKENIDTALLFVSQHL QDILAEMNKTSSSNTVKYDTQHETYMKLAHEILNLGPYFVIQPRSPGSCVFVYKGEVIGRGF NETNCSLSGIRHAELIAIEKILEHYPASVFKETTLYVTVEPCLMCAAALKQLHIKAVYFGCGND RFGGCGSVFSINKDQSIDPSYPVYPGLFYSEAVMLMREFYVQENVKAPVPQSKKQRVLKR EVKSLDLSRFK S. cerevisiae TAD1, SEQ ID NO: 104 MVSCQGTRPCIVNLLTMPSEDKLGEEISTRVINEYSKLKSACRPIIRPSGIREWTILAGVAAIN RDGGANKIEILSIATGVKALPDSELQRSEGKILHDCHAEILALRGANTVLLNRIQNYNPSSGD KFIQHNDEIPARFNLKENWELALYISRLPCGDASMSFLNDNCKNDDFIKIEDSDEFQYVDRS VKTILRGRLNFNRRNVVRTKPGRYDSNITLSKSCSDKLLMKQRSSVLNCLNYELFEKPVFLK YIVIPNLEDETKHHLEQSFHTRLPNLDNEIKFLNCLKPFYDDKLDEEDVPGLMCSVKLFMDDF STEEAILNGVRNGFYTKSSKPLRKHCQSQVSRFAQWELFKKIRPEYEGISYLEFKSRQKKRS QLIIAIKNILSPDGWIPTRTDDVK S. cerevisiae TAD2, SEQ ID NO: 105 MQHIKHMRTAVRLARYALDHDETPVACIFVHTPTGQVMAYGMNDTNKSLTGVAHAEFMGI DQIKAMLGSRGVVDVFKDITLYVTVEPCIMCASALKQLDIGKVVFGCGNERFGGNGTVLSVN HDTCTLVPKNNSAAGYESIPGILRKEAIMLLRYFYVRQNERAPKPRSKSDRVLDKNTFPPME WSKYLNEEAFIETFGDDYRTCFANKVDLSSNSVDWDLIDSHQDNIIQELEEQCKMFKFNVH KKSKV A. thaliana TAD2, SEQ ID NO: 106 MEEDHCEDSHNYMGFALHQAKLALEALEVPVGCVFLEDGKVIASGRNRTNETRNATRHAE MEAIDQLVGQWQKDGLSPSQVAEKFSKCVLYVTCEPCIMCASALSFLGIKEVYYGCPNDKF GGCGSILSLHLGSEEAQRGKGYKCRGGIMAEEAVSLFKCFYEQGNPNAPKPHRPVVQRER T X. laevis ADAT2, SEQ ID NO: 107 MEPLQITEEIQNWMHKAFQMAQDALNNGEVPVGCLMVYGNQWGKGRNEVNETKNATQH AEMVAIDQVLDWCEMNSKKSTDVFENIVLYVTVEPCIMCAGALRLLKIPLWYGCRNERFGG CGSVLNVSGDDIPDTGTKFKCIGGYQAEKAIELLKTFYKQENPNAPKSKVRKKE X. tropicalis ADAT2, SEQ ID NO: 108 MTEEIQNWMHKAFQMAQDALNNGEVPVGCLMVYDNQVVGKGRNEVNETKNATRHAEMV AIDQVLDWCEKNSKKSRDVFENIVLYVTVEPCIMCAGALRLLKIPLWYGCRNERFGGCGSV LNVAGDNIPDTGTEFKYIGGYQAEKAVELLKTFYKQENPNAPRSKVRKKE D. rerio ADAT2, SEQ ID NO: 109 MQEVGVDPEKNDFLQPSDSEVQTWMAKAFDMAVEALENGEVPVGCLMVYNNEIIGKGRN EVNETKNATRHAEMVALDQVLDWCRLREKDCKEVCEQTVLYVTVEPCIMCAAALRLLRIPF VVYGCKNERFGGCGSVLDVSSDHLPHTGTSFKCIAGYRAEEAVEMLKTFYKQENPNAPKP KVRKDSINPQDGAAVIQVMRGPPDEETETIAHLS B. Taurus ADAT2, SEQ ID NO: 110 MEAKAGPTAATDGAYSVSAEETEKWMEQAMQMAKDALDNTEVPVGCLMVYNNEVVGKG RNEVNQTKNATRHAEMVAIDQALDWCRRRGRSPSEVFEHTVLYVTVEPCIMCAAALRLMRI PLVVYGCQNERFGGCGSVLDIASADLPSTGKPFQCTPGYRAEEAVEMLKTFYKQENPNAP KSKVRKKECHKS M. musculus ADAT2, SEQ ID NO: 111 MEEKVESTTTPDGPCVVSVQETEKWMEEAMRMAKEALENIEVPVGCLMVYNNEVVGKGR NEVNQTKNATRHAEMVAIDQVLDWCHQHGQSPSTVFEHTVLYVTVEPCIMCAAALRLMKIP LVVYGCQNERFGGCGSVLNIASADLPNTGRPFQCIPGYRAEEAVELLKTFYKQENPNAPKS KVRKKDCQKS H. sapiens ADAT2 SEQ ID NO: 112 MEAKAAPKPAASGACSVSAEETEKWMEEAMHMAKEALENTEVPVGCLMVYNNEVVGKGR NEVNQTKNATRHAEMVAIDQVLDWCRQSGKSPSEVFEHTVLYVTVEPCIMCAAALRLMKIP LWYGCQNERFGGCGSVLNIASADLPNTGRPFQCIPGYRAEEAVEMLKTFYKQENPNAPKS KVRKKECQKS BE1 for Mammalian expression (rAPOBEC1-XTEN-dCas9-NLS) SEQ ID NO: 113 MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKH VEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHAD PRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCI ILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLKSGSETPGTSESATPESDKK YSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRT ARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYH EKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYN QLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNF DLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSAS MIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKM DGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFR IPYYVGPLARGNSRFAWMTRKSEETITPWNFEEWDKGASAQSFIERMTNFDKNLPNEKVL PKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFK KIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLI HDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENI VIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGR DMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKN YWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKY DENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLES EFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGET GEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKK YGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKD LIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQK QLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGA PAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSPKKKRKV BE2 (rAPOBEC1-XTEN-dCas9-UGI-NLS) SEQ ID NO: 114 MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKH VEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHAD PRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCI ILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLKSGSETPGTSESATPESDKK YSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRT ARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYH EKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYN QLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNF DLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSAS MIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKM DGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFR IPYYVGPLARGNSRFAWMTRKSEETITPWNFEEWDKGASAQSFIERMTNFDKNLPNEKVL PKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFK KIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLI HDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENI VIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGR DMYVDQELDINRLSDYDVDAIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKN YWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKY DENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLES EFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGET GEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKK YGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKD LIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQK QLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGA PAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSTNLSDIIEKETG KQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSN GENKIKMLSGGSPKKKRKV BE3 (rAPOBEC1-XTEN-Cas9n-UGI-NLS) SEQ ID NO: 115 MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKH VEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHAD PRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCI ILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLKSGSETPGTSESATPESDKK YSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRT ARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYH EKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYN QLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNF DLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSAS MIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKM DGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFR IPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVL PKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFK KIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLI HDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENI VIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGR DMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKN YWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKY DENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLES EFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGET GEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKK YGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKD LIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQK QLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGA PAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSTNLSDIIEKETG KQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSN GENKIKMLSGGSPKKKRKV CDA1-BE3: SEQ ID NO: 116 MTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGERRACFWGYAVNKPQSGT ERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKILEWYNQELRGNGHTLKIWA CKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIFIQSSHNQLNENRWLEKTLKR AEKRRSELSIMIQVKILHTTKSPAVSGSETPGTSESATPESDKKYSIGLAIGTNSVGWAVITD EYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIF SNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDK ADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAIL SARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDL DNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALV RQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRK QRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAW MTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTK VKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFN ASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLK RRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVS GQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKN SRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDV DHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDN LTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLV SDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKS EQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPL IETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKK DWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKG YKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSP EDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLF TLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSTNLS DIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWA LVIQDSNGENKIKMLSGGSPKKKRKV AID-BE3: SEQ ID NO: 117 MDSLLMNRRKFLYQFKNVRWAKGRRETYLCYVVKRRDSATSFSLDFGYLRNKNGCHVELL FLRYISDWDLDPGRCYRVTWFTSWSPCYDCARHVADFLRGNPNLSLRIFTARLYFCEDRKA EPEGLRRLHRAGVQIAIMTFKDYFYCWNTFVENHERTFKAWEGLHENSVRLSRQLRRILLP LYEVDDLRDAFRTLGLSGSETPGTSESATPESDKKYSIGLAIGTNSVGWAVITDEYKVPSKK FKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVD DSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLAL AHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAI LSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDD LDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALV RQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRK QRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAW MTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTK VKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFN ASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLK RRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVS GQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKN SRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDV DHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDN LTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLV SDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKS EQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLS MPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKV EKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRM LASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEF SKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTK EVLDATLIHQSITGLYETRIDLSQLGGDSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIG NKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLSGGSPKKKRKV BE3-Gam: SEQ ID NO: 118 MAKPAKRIKSAAAAYVPQNRDAVITDIKRIGDLQREASRLETEMNDAIAEITEKFAARIAPIKT DIETLSKGVQGWCEANRDELTNGGKVKTANLVTGDVSWRVRPPSVSIRGMDAVMETLERL GLQRFIRTKQEINKEAILLEPKAVAGVAGITVKSGIEDFSIIPFEQEAGISGSETPGTSESATPE SSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKH VEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHAD PRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCI ILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLKSGSETPGTSESATPESDKK YSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRT ARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYH EKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYN QLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNF DLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSAS MIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKM DGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFR IPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVL PKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFK KIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEER LKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLI HDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENI VIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGR DMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKN YWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKY DENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLES EFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGET GEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKK YGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKD LIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQK QLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGA PAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSTNLSDIIEKETG KQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSN GENKIKMLSGGSPKKKRKV SaBE3-Gam: SEQ ID NO: 119 MAKPAKRIKSAAAAYVPQNRDAVITDIKRIGDLQREASRLETEMNDAIAEITEKFAARIAPIKT DIETLSKGVQGWCEANRDELTNGGKVKTANLVTGDVSWRVRPPSVSIRGMDAVMETLERL GLQRFIRTKQEINKEAILLEPKAVAGVAGITVKSGIEDFSIIPFEQEAGISGSETPGTSESATPE SSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKH VEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHAD PRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCI ILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLKSGSETPGTSESATPESGKR NYILGLAIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEGRRSKRGARRLKRRRRHRIQR VKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLHLAKRRGVHNVNEVEEDT GNELSTKEQISRNSKALEEKYVAELQLERLKKDGEVRGSINRFKTSDYVKEAKQLLKVQKAY HQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGWKDIKEWYEMLMGHCTYFPEELRSVKYAY NADLYNALNDLNNLVITRDENEKLEYYEKFQIIENVFKQKKKPTLKQIAKEILVNEEDIKGYRV TSTGKPEFTNLKVYHDIKDITARKEIIENAELLDQIAKILTIYQSSEDIQEELTNLNSELTQEEIE QISNLKGYTGTHNLSLKAINLILDELWHTNDNQIAIFNRLKLVPKKVDLSQQKEIPTTLVDDFIL SPVVKRSFIQSIKVINAIIKKYGLPNDIIIELAREKNSKDAQKMINEMQKRNRQTNERIEEIIRTT GKENAKYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVDHIIPRSVSFDNSFNNKVLVK QEENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRISKTKKEYLLEERDINRFSVQK DFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFTSFLRRKWKFKKERNKGYKH HAEDALHANADFIFKEWKKLDKAKKVMENQMFEEKQAESMPEIETEQEYKEIFITPHQIKHIK DFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLNGLYDKDNDKLKKLINKSPEK LLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKYSKKDNGPVIKKIKYYGNKLN AHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKNLDVIKKENYYEVNSKCYEEA KKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLLNRIEVNMIDITYREYLENMNDKRPP RIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQIIKKGGSPKKKRKVSSDYKDHDGDYKDHDI DYKDDDDKSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTD ENVMLLT SDAPEYKPWALVIQDSNGENKIKMLSGGSPKKKRKV BE4: SEQ ID NO: 120 MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKH VEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHAD PRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCI ILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSE SATPESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIG ALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDK KHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLN PDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAIL LSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYID GGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQE DFYPFLKDNREKIEKILTFRIPYYVGPL ARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYE YFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSV EISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLF DDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTF KEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARE NQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQE LDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLN AKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIR EVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYK VYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKG RDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPT VAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYS LFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHK HYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFD TTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSGGSGGSTNLSDIIEKETGK QLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNG ENKIKMLSGGSGGSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYD ESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLSGGSPKKKRK BE4-Gam: SEQ ID NO: 121 MAKPAKRIKSAAAAYVPQNRDAVITDIKRIGDLQREASRLETEMNDAIAEITEKFAARIAPIKT DIETLSKGVQGWCEANRDELTNGGKVKTANLVTGDVSWRVRPPSVSIRGMDAVMETLERL GLQRFIRTKQEINKEAILLEPKAVAGVAGITVKSGIEDFSIIPFEQEAGISGSETPGTSESATPE SSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKH VEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHAD PRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCI ILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSE SATPESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIG ALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDK KHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLN PDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAIL LSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYID GGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQE DFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQ SFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDL LFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDI LEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGK TILDFLKSDGFANRNFMQLIH DDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVI EMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRD MYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNY WRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYD ENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESE FVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETG EIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKY GGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDL HKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQ LFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAP AAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSGGSGGSTNLSDII EKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALV IQDSNGENKIKMLSGGSGGSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDIL VHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLSGGSPKKKRK SaBE4: SEQ ID NO: 122 MSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKH VEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHAD PRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCI ILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSE SATPESSGGSSGGSGKRNYILGLAIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEGRRS KRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLH LAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEVRGSINRFK TSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGWKDIKEWYEML MGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQIIENVFKQKKKPTL KQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEIIENAELLDQIAKILTIYQSSE DIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAINLILDELWHTNDNQIAIFNRLKLVPKK VDLSQQKEIPTTLVDDFILSPVVKRSFIQSIKVINAIIKKYGLPNDIIIELAREKNSKDAQKMINE MQKRNRQTNERIEEIIRTTGKENAKYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVD HIIPRSVSFDNSFNNKVLVKQEENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRIS KTKKEYLLEERDINRFSVQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFT SFLRRKWKFKKERNKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMP EIETEQEYKEIFITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLN GLYDKDNDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKY SKKDNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKN LDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLLNRIEV NMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQIIKKGGSPKKK RKVSSDYKDHDGDYKDHDIDYKDDDDKSGGSGGSGGSTNLSDIIEKETGKQLVIQESILMLP EEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGEN KIKMLSGGSGGSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDES TDENVMLL TSDAPEYKPWALVIQDSNGENKIKMLSGGSPKKKRKV SaBE4-Gam: SEQ ID NO: 123 MAKPAKRIKSAAAAYVPQNRDAVITDIKRIGDLQREASRLETEMNDAIAEITEKFAARIAPIKT DIETLSKGVQGWCEANRDELTNGGKVKTANLVTGDVSWRVRPPSVSIRGMDAVMETLERL GLQRFIRTKQEINKEAILLEPKAVAGVAGITVKSGIEDFSIIPFEQEAGISGSETPGTSESATPE SSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKH VEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHAD PRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCI ILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSE SATPESSGGSSGGSGKRNYILGLAIGITSVGYGIIDYETRDVIDAGVRLFKEANVENNEGRRS KRGARRLKRRRRHRIQRVKKLLFDYNLLTDHSELSGINPYEARVKGLSQKLSEEEFSAALLH LAKRRGVHNVNEVEEDTGNELSTKEQISRNSKALEEKYVAELQLERLKKDGEVRGSINRFK TSDYVKEAKQLLKVQKAYHQLDQSFIDTYIDLLETRRTYYEGPGEGSPFGWKDIKEWYEML MGHCTYFPEELRSVKYAYNADLYNALNDLNNLVITRDENEKLEYYEKFQIIENVFKQKKKPTL KQIAKEILVNEEDIKGYRVTSTGKPEFTNLKVYHDIKDITARKEIIENAELLDQIAKILTIYQSSE DIQEELTNLNSELTQEEIEQISNLKGYTGTHNLSLKAINLILDELWHTNDNQIAIFNRLKLVPKK VDLSQQKEIPTTLVDDFILSPVVKRSFIQSIKVINAIIKKYGLPNDIIIELAREKNSKDAQKMINE MQKRNRQTNERIEEIIRTTGKENAKYLIEKIKLHDMQEGKCLYSLEAIPLEDLLNNPFNYEVD HIIPRSVSFDNSFNNKVLVKQEENSKKGNRTPFQYLSSSDSKISYETFKKHILNLAKGKGRIS KTKKEYLLEERDINRFSVQKDFINRNLVDTRYATRGLMNLLRSYFRVNNLDVKVKSINGGFT SFLRRKWKFKKERNKGYKHHAEDALIIANADFIFKEWKKLDKAKKVMENQMFEEKQAESMP EIETEQEYKEIFITPHQIKHIKDFKDYKYSHRVDKKPNRELINDTLYSTRKDDKGNTLIVNNLN GLYDKDNDKLKKLINKSPEKLLMYHHDPQTYQKLKLIMEQYGDEKNPLYKYYEETGNYLTKY SKKDNGPVIKKIKYYGNKLNAHLDITDDYPNSRNKVVKLSLKPYRFDVYLDNGVYKFVTVKN LDVIKKENYYEVNSKCYEEAKKLKKISNQAEFIASFYNNDLIKINGELYRVIGVNNDLLNRIEV NMIDITYREYLENMNDKRPPRIIKTIASKTQSIKKYSTDILGNLYEVKSKKHPQIIKKGGSPKKK RKVSSDYKDHDGDYKDHDIDYKDDDDKSGGSGGSGGSTNLSDIIEKETGKQLVIQESILMLP EEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLSGGS GGSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLT SDAPEYKPWALVIQDSNGENKIKMLSGGSPKKKRKV BE4max and AncBE4max, SEQ ID NO: 124 MKRTADGSEFESPKKKRKV[APOBEC or ancestral APOBEC, sequences see below]SGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEY KVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSN EMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKAD LRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSA RLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLD NLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQ RTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWM TRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNA SLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKR RRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSG QGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNS RERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVD HIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNL TKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLV SDFRKDFQFYKVREINNYHHAHDAYLNAWGTALIKKYPKLESEFVYGDYKVYDVRKMIAKS EQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLS MPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKV EKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRM LASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEF SKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTK EVLDATLIHQSITGLYETRIDLSQLGGDSGGSGGSGGSTNLSDIIEKETGKQLVIQESILMLPE EVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKML_SGGS GGSGGS_TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLL TSDAPEYKPWALVIQDSNGENKIKMLSGGSKRTADGSEFEPKKKRKV Rat APOBEC1, SEQ ID NO: 125 SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKHV EVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHADP RNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIIL GLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLK Anc689 APOBEC, SEQ ID NO: 126 SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEIKWGTSHKIWRHSSKNTTKHVE VNFIEKFTSERHFCPSTSCSITWFLSWSPCGECSKAITEFLSQHPNVTLVIYVARLYHHMDQ QNRQGLRDLVNSGVTIQIMTAPEYDYCWRNFVNYPPGKEAHWPRYPPLWMKLYALELHA GILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWAT GLK Anc687 APOBEC, SEQ ID NO: 127 SSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKEACLLYEIKWGTSHKIWRNSGKNTTKHVE VNFIEKFTSERHFCPSISCSITWFLSWSPCWECSKAIREFLSQHPNVTLVIYVARLFQHMDQ QNRQGLRDLVNSGVTIQIMTASEYDHCWRNFVNYPPGKEAHWPRYPPLWMKLYALELHA GILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLK Anc686 APOBEC, SEQ ID NO: 128 SSETGPVAVDPTLRRRIEPEFFNRNYDPRELRKETYLLYEIKWGKESKIWRHTSNNRTQHA EVNFLENFFNELYFNPSTHCSITWFLSWSPCGECSKAIVEFLKEHPNVNLEIYVARLYLCED ERNRQGLRDLVNSGVTIRIMNLPDYNYCWRTFVSHQGGDEDYWPRHFAPWVRLYVLELY CIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWAT GLK Anc655 APOBEC, SEQ ID NO: 129 SSETGPVAVDPTLRRRIEPFYFQFNNDPRACRRKTYLCYELKQDGSTWVWKRTLHNKGRH AEICFLEKISSLEKLDPAQHYRITWYMSWSPCSNCAQKIVDFLKEHPHVNLRIYVARLYYHEE ERYQEGLRNLRRSGVSIRVMDLPDFEHCWETFVDNGGGPFQPWPGLEELNSKQLSRRLQ AGILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHIL WATGLK Anc733 APOBEC, SEQ ID NO: 130 SSETGPVAVDPTLRRRIEPFHFQFNNDPRAYRRKTYLCYELKQDGSTWVLDRTLRNKGRH AEICFLDKINSWERLDPAQHYRVTWYMSWSPCSNCAQQVVDFLKEHPHVNLRIFAARLYYH EQRRYQEGLRSLRGSGVPVAVMTLPDFEHCWETFVDHGGRPFQPWDGLEELNSRSLSRR LQAGILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHI LWATGLK APOBEC ancestor #686, SEQ ID NO: 131 EFFNRNYDPRELRKETYLLYEIKWGKESKIWRHWCTSNNRTQHAEVNFLENFFNELYFNPS THCSITWFLSWSPCGECSKAIVEFLKEHPNVNLEIYVARLYLCEDERNRQGLRDLVNSGVTI RIMNLPDYNYCWRTFVSHQGGDEDYWPRHFAPWVRL APOBEC ancestor #733, SEQ ID NO: 132 FHFQFNNDPRAYRRKTYLCYELKQDGSTWVLDRGCTLRNKGRHAEICFLDKINSWERLDP AQHYRVTWYMSWSPCSNCAQQVVDFLKEHPHVNLRIFAARLYYHEQRRYQEGLRSLRGS GVPVAVMTLPDFEHCWETFVDHGGRPFQPWDGLEELNSRSLSRRLQAG APOBEC ancestor #656_FERNY, SEQ ID NO: 133 FERNYDPRELRKETYLLYEIKWGKSGKLWRHWCQNNRTQHAEVYFLENIFNARRFNPSTH CSITWYLSWSPCAECSQKIVDFLKEHPNVNLEIYVARLYYHEDERNRQGLRDLVNSGVTIRI MDLPDYNYCWKTFVSDQGGDEDYWPGHFAPWIKQYSLKL APOBEC ancestor #655, SEQ ID NO: 134 FYFQFNNDPRACRRKTYLCYELKQDGSTWVWKRGCTLHNKGRHAEICFLEKISSLEKLDPA QHYRITWYMSWSPCSNCAQKIVDFLKEHPHVNLRIYVARLYYHEEERYQEGLRNLRRSGV SIRVMDLPDFEHCWETFVDNGGGPFQPWPGLEENSKQLSRRLQAG APOBEC ancestor #649, SEQ ID NO: 135 FYEEFNNTLKSCRHKTLLCFSLKQDENTTLWKWGYAHNNGRHAEILVLREIENYEKLDPAA KYRITLYMSYSPCNDCADKIVDFLKKHPNVNLNIKVSRLYYHEDEKYQEGLRNLKQPGVSLK VMDRSDFEECFDLFVDPGGGEFQPWPGLEEKSKQYSATLQAG ABE6.3, SED ID: 136 MSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGWNRPIGRHDPTAHAEI MALRQGGLVMQNYRLIDATYVTLEPCVMCAGAMIHSRIGRVVFGARDAKTGAAGSLMDVL HHPGMNHRVEITEGILADECAALLSDFFRMRRQEIKAQKKAQSSTDSGGSSGGSSGSETP GTSESATPESSGGSSGGSSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVLNNRVIGE GWNRSIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVF GVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALLCYFFRMRRQVFNAQKKAQS STDSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYKV PSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEM AKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRL IYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLS KSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLA QIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLP EKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFD NGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKS EETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVT EGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGT YHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYT GWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDS LHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERM KRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVP QSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAE RGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFR KDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEI GKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQV NIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKS KKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAG ELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVI LADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDA TLIHQSITGLYETRIDLSQLGGDSGGSPKKKRKV ABE7.8, SED ID: 137 MSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGWNRPIGRHDPTAHAEI MALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSRIGRVVFGARDAKTGAAGSLMDVL HHPGMNHRVEITEGILADECAALLSDFFRMRRQEIKAQKKAQSSTDSGGSSGGSSGSETP GTSESATPESSGGSSGGSSEVEFSHEYWMRHALTLAKRALDEREVPVGAVLVLNNRVIGE GWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVF GVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECNALLCYFFRMRRQVFNAQKKAQS STDSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYKV PSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEM AKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRL IYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLS KSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLA QIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLP EKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFD NGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKS EETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVT EGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGT YHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYT GWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDS LHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERM KRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVP QSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAE RGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFR KDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEI GKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQV NIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKS KKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAG ELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVI LADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDA TLIHQSITGLYETRIDLSQLGGDSGGSPKKKRKV ABE7.9, SED ID: 138 MSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGWNRPIGRHDPTAHAEI MALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSRIGRVVFGARDAKTGAAGSLMDVL HHPGMNHRVEITEGILADECAALLSDFFRMRRQEIKAQKKAQSSTDSGGSSGGSSGSETP GTSESATPESSGGSSGGSSEVEFSHEYWMRHALTLAKRALDEREVPVGAVLVLNNRVIGE GWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVF GVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECNALLCYFFRMPRQVFNAQKKAQS STDSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYKV PSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEM AKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRL IYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLS KSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLA QIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLP EKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFD NGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKS EETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVT EGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGT YHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYT GWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDS LHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERM KRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVP QSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAE RGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFR KDFQFYKVREINNYHHAHDAYLNAWGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEI GKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQV NIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKS KKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAG ELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVI LADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDA TLIHQSITGLYETRIDLSQLGGDSGGSPKKKRKV ABE7.10, SED ID: 139 MSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGWNRPIGRHDPTAHAEI MALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSRIGRVVFGARDAKTGAAGSLMDVL HHPGMNHRVEITEGILADECAALLSDFFRMRRQEIKAQKKAQSSTDSGGSSGGSSGSETP GTSESATPESSGGSSGGSSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGE GWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVF GVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALLCYFFRMPRQVFNAQKKAQS STDSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYKV PSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEM AKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRL IYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLS KSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLA QIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLP EKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFD NGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKS EETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVT EGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGT YHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYT GWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDS LHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERM KRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVP QSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAE RGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFR KDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEI GKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQV NIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKS KKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAG ELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVI LADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDA TLIHQSITGLYETRIDLSQLGGDSGGSPKKKRKV ABEmax, SEQ ID NO: 140 MKRTADGSEFESPKKKRKVSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIG EGWNRPIGRHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSRIGRW FGARDAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLSDFFRMRRQEIKAQKKAQS STDSGGSSGGSSGSETPGTSESATPESSGGSSGGSSEVEFSHEYWMRHALTLAKRARDE REVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEP CVMCAGAMIHSRIGRVVFGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALLCY FFRMPRQVFNAQKKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYSIG LAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRR YTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYP TIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFE ENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAE DAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKR YDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGT EELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPY YVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPK HSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIE CFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKT YAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHD DSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIE MARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDM YVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYW RQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDE NDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEF VYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEI VWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGG FDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIK LPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLF VEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAA FKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKK RKV SpACE, SEQ ID NO: 140 MKRTADGSEFESPKKKRKVSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIG EGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYGTFEPCVMCAGAMIHSRIGRVV FGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALLCYFFRMPRQVFNAQKKAQ SSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYK VPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNE MAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADL RLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSAR LSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNL LAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQ LPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTF DNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRK SEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYV TEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLG TYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRY TGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGD SLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRER MKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIV PQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKA ERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDF RKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQE IGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQ VNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGK SKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASA GELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKR VILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKKRKVSGGSSGGSSGSETPG TSESATPESSGGSSGGSTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGER RACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKILE WYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIFIQS SHNQLNENRWLEKTLKRAEKWRSELSIMIQVKILHTTKSPAVSGGSGGSGGSTNLSDIIEKE TGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQD SNGENKIKMLSGGSGGSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVH TAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLGSGATNFSLLKQAGDVEENPG PMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPT LVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTL VNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHY QQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKSGGSP KKKRK SPACEΔUGI, SEQ ID NO: 141 MKRTADGSEFESPKKKRKVSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIG EGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYGTFEPCVMCAGAMIHSRIGRVV FGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALLCYFFRMPRQVFNAQKKAQ SSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYK VPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNE MAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADL RLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSAR LSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNL LAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQ LPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTF DNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRK SEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYV TEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLG TYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRY TGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGD SLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRER MKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIV PQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKA ERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDF RKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQE IGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQ VNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGK SKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASA GELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKR VILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVL DATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKKRKVSGGSSGGSSGSETPG TSESATPESSGGSSGGSTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGER RACFWGYAVN KPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKILE WYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIFIQS SHNQLNENRWLEKTLKRAEKWRSELSIMIQVKILHTTKSPAVGSGATNFSLLKQAGDVEEN PGPMVSKGEELFTGWPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPW PTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGD TLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLAD HYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKSGG SPKKKRKV SPACE-NG, SEQ ID NO: 142 MKRTADGSEFESPKKKRKVSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIG EGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYGTFEPCVMCAGAMIHSRIGRVV FGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALLCYFFRMPRQVFNAQKKAQ SSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYK VPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNE MAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADL RLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSAR LSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNL LAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQ LPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTF DNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRK SEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYV TEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLG TYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRY TGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGD SLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRER MKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIV PQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKA ERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDF RKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQE IGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQ VNIVKKTEVQTGGFSKESIRPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKG KSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLAS ARFLQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSK RVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPRAFKYFDTTIDRKVYRSTKEV LDATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKKRKVSGGSSGGSSGSETP GTSESATPESSGGSSGGSTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGE RRACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKIL EWYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIFIQ SSHNQLNENRWLEKTLKRAEKWRSELSIMIQVKILHTTKSPAVSGGSGGSGGSTNLSDIIEK ETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQ DSNGENKIKMLSGGSGGSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILV HTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLGSGATNFSLLKQAGDVEENP GPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWP TLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTL VNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHY QQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKSGGSP KKKRKV SPACE-VRQR, SEQ ID NO: 143 MKRTADGSEFESPKKKRKVSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIG EGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYGTFEPCVMCAGAMIHSRIGRVV FGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALLCYFFRMPRQVFNAQKKAQ SSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYK VPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNE MAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADL RLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSAR LSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNL LAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQ LPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTF DNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRK SEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYV TEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLG TYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRY TGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGD SLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRER MKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIV PQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKA ERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDF RKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQE IGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQ VNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGK SKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASA RELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKR VILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKQYRSTKEVL DATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKKRKVSGGSSGGSSGSETPG TSESATPESSGGSSGGSTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGER RACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKILE WYNQELRGNGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIFIQS SHNQLNENRWLEKTLKRAEKWRSELSIMIQVKILHTTKSPAVSGGSGGSGGSTNLSDIIEKE TGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQD SNGENKIKMLSGGSGGSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVH TAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLGSGATNFSLLKQAGDVEENPG PMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPT LVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTL VNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHY QQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKSGGSP KKKRKV SPACE-NAA, SEQ ID NO: 144 MKRTADGSEFESPKKKRKVSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIG EGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYGTFEPCVMCAGAMIHSRIGRVV FGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALLCYFFRMPRQVFNAQKKAQ SSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYK VPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNE MAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADL RLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSAR LSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNL LAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQ LPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTF DNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRK SEETITPWNFEEWDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYV TEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLG TYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRY TGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGD SLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENMEMARENQTTQKGQKNSRER MKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIV PQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKA ERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDF RKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQE IGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQ VNIVKKTEIQTVGQNGGLFDDNPKSPLEVTPSKLVPLKKELNPKKYGGYQKPTTAYPVLLITD TKQLIPISVMNKKQFEQNPVKFLRDRGYQQVGKNDFIKLPKYTLVDIGDGIKRLWASSKEIHK GNQLVVSKKSQILLYHAHHLDSDLSNDYLQNHNQQFDVLFNEIISFSKKCKLGKEHIQKIENV YSNKKNSASIEELAESFIKLLGFTQLGATSPFNFLGVKLNQKQYKGKKDYILPCTEGTLIRQSI TGLYETRVDLSKIGEDSGGSKRTADGSEFEPKKKRKVSGGSSGGSSGSETPGTSESATPE SSGGSSGGSTDAEYVRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGERRACFWGYA VNKPQSGTERGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKILEWYNQELRG NGHTLKIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIFIQSSHNQLNEN RWLEKTLKRAEKWRSELSIMIQVKILHTTKSPAVSGGSGGSGGSTNLSDIIEKETGKQLVIQE SILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKM LSGGSGGSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDE NVMLLTSDAPEYKPWALVIQDSNGENKIKMLGSGATNFSLLKQAGDVEENPGPMVSKGEE LFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGV QCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGID FKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDG PVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKSGGSPKKKRKV BE4max-ΔUG1-eUNG, SEQ ID NO: 147 MKRTADGSEFESPKKKRKVSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEIN WGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFL SRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPSNEAH WPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPPHILWATGLK SGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYKVPSK KFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKV DDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYL ALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKS RRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQI GDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEK YKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNG SIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEE TITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEG MRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYH DLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGW GRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLH EHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKR IEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSF LKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGG LSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDF QFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKA TAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIV KKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKK LKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGEL QKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILA DANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATL IHQSITGLYETRIDLSQLGGDSGGSGGSGGSANELTWHDVLAEEKQQPYFLNTLQTVASER QSGVTIYPPQKDVFNAFRFTELGDVKVVILGQDPYHGPGQAHGLAFSVRPGIAIPPSLLNMY KELENTIPGFTRPNHGYLESWARQGVLLLNTVLTVRAGQAHSHASLGWETFTDKVISLINQH REGVVFLLWGSHAQKKGAIIDKQRHHVLKAPHPSPLSAHRGFFGCNHFVLANQWLEQRGE TPIDWMPVLPAESESGGSKRTADGSEFEPKKKRKVGGGGSGATNFSLLKQAGDVEENPG PMVSKGEELFTGWPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPT LVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTL VNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHY QQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK BE4max(R33A)-ΔUGI-eUNG, SEQ ID NO: 148 MKRTADGSEFESPKKKRKVSSETGPVAVDPTLRRRIEPHEFEVFFDPRELAKETCLL YEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGE CSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCW RNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQ SCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYS IGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRL KRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNL SDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQS KNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSE ETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVE DRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFD DKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDD SLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLY YLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQI TKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIM NFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQ TGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLWAKVEKGKSKKL KSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLAS AGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQI SEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTID RKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSGGSGGSANELTWHDVL AEEKQQPYFLNTLQTVASERQSGVTIYPPQKDVFNAFRFTELGDVKVVILGQDPYH GPGQAHGLAFSVRPGIAIPPSLLNMYKELENTIPGFTRPNHGYLESWARQGVLLLNT VLTVRAGQAHSHASLGWETFTDKVISLINQHREGVVFLLWGSHAQKKGAIIDKQRH HVLKAPHPSPLSAHRGFFGCNHFVLANQWLEQRGETPIDWMPVLPAESESGGSKR TADGSEFEPKKKRKVGGGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPI LVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQC FSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELK GIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQ QNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK BE4max(R33A/K34A)-ΔUGI-eUNG, SEQ ID NO: 149 MKRTADGSEFESPKKKRKVSSETGPVAVDPTLRRRIEPHEFEVFFDPRELAAETCLL YEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGE CSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCW RNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQ SCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYS IGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRL KRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNL SDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQS KNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSE ETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVE DRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFD DKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDD SLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLY YLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQI TKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIM NFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQ TGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLWAKVEKGKSKKL KSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLAS AGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQI SEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTID RKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSGGSGGSANELTWHDVL AEEKQQPYFLNTLQTVASERQSGVTIYPPQKDVFNAFRFTELGDVKVVILGQDPYH GPGQAHGLAFSVRPGIAIPPSLLNMYKELENTIPGFTRPNHGYLESWARQGVLLLNT VLTVRAGQAHSHASLGWETFTDKVISLINQHREGVVFLLWGSHAQKKGAIIDKQRH HVLKAPHPSPLSAHRGFFGCNHFVLANQWLEQRGETPIDWMPVLPAESESGGSKR TADGSEFEPKKKRKVGGGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPI LVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQC FSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELK GIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQ QNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK nCas9-eUNG for BE4max, SEQ ID NO: 150 MKRTADGSEFESPKKKRKVSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKK YSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEAT RLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIF GNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDN SDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNG LFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAK NLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFD QSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSI PHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRK SEETITPWNFEEWDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELT KVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISG VEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHL FDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIH DDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHK PENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSD NVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVET RQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYH HAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFY SNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKK TEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGK SKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKR MLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDE IIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFD TTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSGGSGGSANELTWH DVLAEEKQQPYFLNTLQTVASERQSGVTIYPPQKDVFNAFRFTELGDVKVVILGQDP YHGPGQAHGLAFSVRPGIAIPPSLLNMYKELENTIPGFTRPNHGYLESWARQGVLLL NTVLTVRAGQAHSHASLGWETFTDKVISLINQHREGVVFLLWGSHAQKKGAIIDKQR HHVLKAPHPSPLSAHRGFFGCNHFVLANQWLEQRGETPIDWMPVLPAESESGGSK RTADGSEFEPKKKRKVGGGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVP ILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQC FSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELK GIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQ QNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK eUNG-BE4max-ΔUGI, SEQ ID NO: 151 MKRTADGSEFESPKKKRKVANELTWHDVLAEEKQQPYFLNTLQTVASERQSGVTIY PPQKDVFNAFRFTELGDVKVVILGQDPYHGPGQAHGLAFSVRPGIAIPPSLLNMYKE LENTIPGFTRPNHGYLESWARQGVLLLNTVLTVRAGQAHSHASLGWETFTDKVISLI NQHREGVVFLLWGSHAQKKGAIIDKQRHHVLKAPHPSPLSAHRGFFGCNHFVLAN QWLEQRGETPIDWMPVLPAESESGGSGGSGGSSSETGPVAVDPTLRRRIEPHEFE VFFDPRELRKETCLLYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTR CSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGV TIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNIL RRKQPQLTFFTIALQSCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSESATPE SSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIG ALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFL VEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFR GHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDL LRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLAR GNSRFAWMTRKSEETITPWNFEEWDKGASAQSFIERMTNFDKNLPNEKVLPKHSL LYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFK KIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDRE MIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDG FANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVD ELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVE NTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTR SDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKA GFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQF YKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEI GKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVL SMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSV LVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSL FELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFV EQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLG APAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTAD GSEFEPKKKRKVGGGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGWPILVEL DGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRY PDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDF KEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTP IGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK eUNG-BE4max(R33A)-ΔUGI = CGBE1, SEQ ID NO: 152 MKRTADGSEFESPKKKRKVANELTWHDVLAEEKQQPYFLNTLQTVASERQSGVTIY PPQKDVFNAFRFTELGDVKVVILGQDPYHGPGQAHGLAFSVRPGIAIPPSLLNMYKE LENTIPGFTRPNHGYLESWARQGVLLLNTVLTVRAGQAHSHASLGWETFTDKVISLI NQHREGVVFLLWGSHAQKKGAIIDKQRHHVLKAPHPSPLSAHRGFFGCNHFVLAN QWLEQRGETPIDWMPVLPAESESGGSGGSGGSSSETGPVAVDPTLRRRIEPHEFE VFFDPRELAKETCLLYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTR CSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGV TIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNIL RRKQPQLTFFTIALQSCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSESATPE SSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIG ALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFL VEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFR GHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDL LRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLAR GNSRFAWMTRKSEETITPWNFEEWDKGASAQSFIERMTNFDKNLPNEKVLPKHSL LYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFK KIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDRE MIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDG FANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVD ELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVE NTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTR SDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKA GFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQF YKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEI GKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVL SMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSV LVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSL FELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFV EQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLG APAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTAD GSEFEPKKKRKVGGGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGWPILVEL DGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRY PDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDF KEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTP IGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK eUNG-BE4max(R33A/K34A)-ΔUGI, SEQ ID NO: 153 MKRTADGSEFESPKKKRKVANELTWHDVLAEEKQQPYFLNTLQTVASERQSGVTIY PPQKDVFNAFRFTELGDVKVVILGQDPYHGPGQAHGLAFSVRPGIAIPPSLLNMYKE LENTIPGFTRPNHGYLESWARQGVLLLNTVLTVRAGQAHSHASLGWETFTDKVISLI NQHREGVVFLLWGSHAQKKGAIIDKQRHHVLKAPHPSPLSAHRGFFGCNHFVLAN QWLEQRGETPIDWMPVLPAESESGGSGGSGGSSSETGPVAVDPTLRRRIEPHEFE VFFDPRELAAETCLLYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTR CSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGV TIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNIL RRKQPQLTFFTIALQSCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSESATPE SSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIG ALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFL VEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFR GHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDL LRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLAR GNSRFAWMTRKSEETITPWNFEEWDKGASAQSFIERMTNFDKNLPNEKVLPKHSL LYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFK KIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDRE MIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDG FANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVD ELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVE NTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTR SDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKA GFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQF YKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEI GKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVL SMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSV LVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSL FELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFV EQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLG APAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTAD GSEFEPKKKRKVGGGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGWPILVEL DGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRY PDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDF KEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTP IGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK eUNG-nCas9 for BE4max, SEQ ID NO: 154 MKRTADGSEFESPKKKRKVANELTWHDVLAEEKQQPYFLNTLQTVASERQSGVTIY PPQKDVFNAFRFTELGDVKVVILGQDPYHGPGQAHGLAFSVRPGIAIPPSLLNMYKE LENTIPGFTRPNHGYLESWARQGVLLLNTVLTVRAGQAHSHASLGWETFTDKVISLI NQHREGVVFLLWGSHAQKKGAIIDKQRHHVLKAPHPSPLSAHRGFFGCNHFVLAN QWLEQRGETPIDWMPVLPAESESGGSGGSGGSSGGSSGGSSGSETPGTSESATP ESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLI GALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLA QIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALV RQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNRE DLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPL ARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPK HSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKED YFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFE DREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLK SDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTV KWDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKE HPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDN KVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLS ELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRK DFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKS EQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATV RKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTV AYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKL PKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQ KQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFT LTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGS KRTADGSEFEPKKKRKVGGGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVV PILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQ CFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIEL KGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHY QQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK BE4max-ΔUGI-hUNG, SEQ ID NO: 155 MKRTADGSEFESPKKKRKVSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCL LYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCG ECSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYC WRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIAL QSCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKY SIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATR LKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFG NIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNS DVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGL FGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKN LSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQ SKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIP HQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKS EETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTK VKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGV EDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLF DDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHD DSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKP ENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDN VPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETR QITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHH AHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYS NIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKT EVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKS KKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRM LASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEII EQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDT TIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSGGSGGSIGQKTLYSF FSPSPARKRHAPSPEPAVQGTGVAGVPEESGDAAAIPAKKAPAGQEEPGTPPSSP LSAEQLDRIQRNKAAALLRLAARNVPVGFGESWKKHLSGEFGKPYFIKLMGFVAEE RKHYTVYPPPHQVFTWTQMCDIKDVKVVILGQDPYHGPNQAHGLCFSVQRPVPPP PSLENIYKELSTDIEDFVHPGHGDLSGWAKQGVLLLNAVLTVRAHQANSHKERGWE QFTDAVVSWLNQNSNGLVFLLWGSYAQKKGSAIDRKRHHVLQTAHPSPLSVYRGF FGCRHFSKTNELLQKSGKKPIDWKELSGGSKRTADGSEFEPKKKRKVGGGGSGAT NFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDA TYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYV QERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNV YIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSAL SKDPNEKRDHMVLLEFVTAAGITLGMDELYK BE4max(R33A)-ΔUGI-hUNG, SEQ ID NO: 156 MKRTADGSEFESPKKKRKVSSETGPVAVDPTLRRRIEPHEFEVFFDPRELAKETCLL YEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGE CSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCW RNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQ SCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYS IGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRL KRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNL SDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQS KNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSE ETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVE DRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFD DKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDD SLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLY YLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQI TKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIM NFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQ TGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKL KSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLAS AGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQI SEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTID RKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSGGSGGSIGQKTLYSFFS PSPARKRHAPSPEPAVQGTGVAGVPEESGDAAAIPAKKAPAGQEEPGTPPSSPLS AEQLDRIQRNKAAALLRLAARNVPVGFGESWKKHLSGEFGKPYFIKLMGFVAEERK HYTVYPPPHQVFTWTQMCDIKDVKVVILGQDPYHGPNQAHGLCFSVQRPVPPPPS LENIYKELSTDIEDFVHPGHGDLSGWAKQGVLLLNAVLTVRAHQANSHKERGWEQF TDAVVSWLNQNSNGLVFLLWGSYAQKKGSAIDRKRHHVLQTAHPSPLSVYRGFFG CRHFSKTNELLQKSGKKPIDWKELSGGSKRTADGSEFEPKKKRKVGGGGSGATNF SLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATY GKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQE RTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIM ADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKD PNEKRDHMVLLEFVTAAGITLGMDELYK BE4max(R33A/K34A)-ΔUGI-hUNG, SEQ ID NO: 157 MKRTADGSEFESPKKKRKVSSETGPVAVDPTLRRRIEPHEFEVFFDPRELAAETCLL YEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGE CSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCW RNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQ SCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYS IGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRL KRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNL SDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQS KNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSE ETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVE DRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFD DKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDD SLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLY YLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQI TKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIM NFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQ TGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKL KSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLAS AGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQI SEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTID RKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSGGSGGSIGQKTLYSFFS PSPARKRHAPSPEPAVQGTGVAGVPEESGDAAAIPAKKAPAGQEEPGTPPSSPLS AEQLDRIQRNKAAALLRLAARNVPVGFGESWKKHLSGEFGKPYFIKLMGFVAEERK HYTVYPPPHQVFTWTQMCDIKDVKVVILGQDPYHGPNQAHGLCFSVQRPVPPPPS LENIYKELSTDIEDFVHPGHGDLSGWAKQGVLLLNAVLTVRAHQANSHKERGWEQF TDAVVSWLNQNSNGLVFLLWGSYAQKKGSAIDRKRHHVLQTAHPSPLSVYRGFFG CRHFSKTNELLQKSGKKPIDWKELSGGSKRTADGSEFEPKKKRKVGGGGSGATNF SLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATY GKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQE RTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIM ADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKD PNEKRDHMVLLEFVTAAGITLGMDELYK nCas9-hUNG for BE4max, SEQ ID NO: 158 MKRTADGSEFESPKKKRKVSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKK YSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEAT RLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIF GNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDN SDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNG LFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAK NLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFD QSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSI PHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRK SEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELT KVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISG VEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHL FDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIH DDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHK PENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSD NVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVET RQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYH HAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFY SNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKK TEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGK SKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKR MLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDE IIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFD TTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSGGSGGSIGQKTLYS FFSPSPARKRHAPSPEPAVQGTGVAGVPEESGDAAAIPAKKAPAGQEEPGTPPSS PLSAEQLDRIQRNKAAALLRLAARNVPVGFGESWKKHLSGEFGKPYFIKLMGFVAE ERKHYTVYPPPHQVFTWTQMCDIKDVKVVILGQDPYHGPNQAHGLCFSVQRPVPP PPSLENIYKELSTDIEDFVHPGHGDLSGWAKQGVLLLNAVLTVRAHQANSHKERGW EQFTDAVVSWLNQNSNGLVFLLWGSYAQKKGSAIDRKRHHVLQTAHPSPLSVYRG FFGCRHFSKTNELLQKSGKKPIDWKELSGGSKRTADGSEFEPKKKRKVGGGGSGA TNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGD ATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGY VQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN VYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSA LSKDPNEKRDHMVLLEFVTAAGITLGMDELYK hUNG-BE4max-ΔUGI, SEQ ID NO: 159 MKRTADGSEFESPKKKRKVIGQKTLYSFFSPSPARKRHAPSPEPAVQGTGVAGVP EESGDAAAIPAKKAPAGQEEPGTPPSSPLSAEQLDRIQRNKAAALLRLAARNVPVG FGESWKKHLSGEFGKPYFIKLMGFVAEERKHYTVYPPPHQVFTWTQMCDIKDVKV VILGQDPYHGPNQAHGLCFSVQRPVPPPPSLENIYKELSTDIEDFVHPGHGDLSGW AKQGVLLLNAVLTVRAHQANSHKERGWEQFTDAVVSWLNQNSNGLVFLLWGSYA QKKGSAIDRKRHHVLQTAHPSPLSVYRGFFGCRHFSKTNELLQKSGKKPIDWKELS GGSGGSGGSSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRH SIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPS NEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPP HILWATGLKSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNSV GWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYT RRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHE KYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLV QTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLG LTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDIL RVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYID GGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAI LRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFE EVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMR KPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGT YHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLK RRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQ KAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARE NQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDM YVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKK MKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQIL DSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAV VGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEIT LANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLG ITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGN ELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILA DANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKE VLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKKRKVGGGGSGAT NFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDA TYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYV QERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNV YIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSAL SKDPNEKRDHMVLLEFVTAAGITLGMDELYK hUNG-BE4max(R33A)-ΔUGI, SEQ ID NO: 160 MKRTADGSEFESPKKKRKVIGQKTLYSFFSPSPARKRHAPSPEPAVQGTGVAGVP EESGDAAAIPAKKAPAGQEEPGTPPSSPLSAEQLDRIQRNKAAALLRLAARNVPVG FGESWKKHLSGEFGKPYFIKLMGFVAEERKHYTVYPPPHQVFTWTQMCDIKDVKV VILGQDPYHGPNQAHGLCFSVQRPVPPPPSLENIYKELSTDIEDFVHPGHGDLSGW AKQGVLLLNAVLTVRAHQANSHKERGWEQFTDAVVSWLNQNSNGLVFLLWGSYA QKKGSAIDRKRHHVLQTAHPSPLSVYRGFFGCRHFSKTNELLQKSGKKPIDWKELS GGSGGSGGSSSETGPVAVDPTLRRRIEPHEFEVFFDPRELAKETCLLYEINWGGRH SIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPS NEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPP HILWATGLKSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNSV GWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYT RRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHE KYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLV QTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLG LTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDIL RVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYID GGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAI LRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFE EVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMR KPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGT YHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLK RRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQ KAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARE NQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDM YVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKK MKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQIL DSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAV VGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEIT LANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLG ITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGN ELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILA DANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKE VLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKKRKVGGGGSGAT NFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDA TYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYV QERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNV YIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSAL SKDPNEKRDHMVLLEFVTAAGITLGMDELYK hUNG-BE4max(R33A/K34A)-ΔUGI, SEQ ID NO: 161 MKRTADGSEFESPKKKRKVIGQKTLYSFFSPSPARKRHAPSPEPAVQGTGVAGVP EESGDAAAIPAKKAPAGQEEPGTPPSSPLSAEQLDRIQRNKAAALLRLAARNVPVG FGESWKKHLSGEFGKPYFIKLMGFVAEERKHYTVYPPPHQVFTWTQMCDIKDVKV VILGQDPYHGPNQAHGLCFSVQRPVPPPPSLENIYKELSTDIEDFVHPGHGDLSGW AKQGVLLLNAVLTVRAHQANSHKERGWEQFTDAVVSWLNQNSNGLVFLLWGSYA QKKGSAIDRKRHHVLQTAHPSPLSVYRGFFGCRHFSKTNELLQKSGKKPIDWKELS GGSGGSGGSSSETGPVAVDPTLRRRIEPHEFEVFFDPRELAAETCLLYEINWGGRH SIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLS RYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCWRNFVNYSPS NEAHWPRYPHLWRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQSCHYQRLPP HILWATGLKSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNSV GWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYT RRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHE KYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLV QTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLG LTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDIL RVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYID GGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAI LRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFE EVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMR KPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGT YHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLK RRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQ KAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARE NQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDM YVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKK MKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQIL DSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAV VGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEIT LANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLG ITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGN ELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILA DANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKE VLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKKRKVGGGGSGAT NFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDA TYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYV QERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNV YIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSAL SKDPNEKRDHMVLLEFVTAAGITLGMDELYK hUNG-nCas9 for BE4max, SEQ ID NO: 162 MKRTADGSEFESPKKKRKVIGQKTLYSFFSPSPARKRHAPSPEPAVQGTGVAGVP EESGDAAAIPAKKAPAGQEEPGTPPSSPLSAEQLDRIQRNKAAALLRLAARNVPVG FGESWKKHLSGEFGKPYFIKLMGFVAEERKHYTVYPPPHQVFTWTQMCDIKDVKV VILGQDPYHGPNQAHGLCFSVQRPVPPPPSLENIYKELSTDIEDFVHPGHGDLSGW AKQGVLLLNAVLTVRAHQANSHKERGWEQFTDAVVSWLNQNSNGLVFLLWGSYA QKKGSAIDRKRHHVLQTAHPSPLSVYRGFFGCRHFSKTNELLQKSGKKPIDWKELS GGSGGSGGSSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNS VGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRY TRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYH EKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQL VQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSL GLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSD ILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYI DGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHA ILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFE EVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMR KPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGT YHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLK RRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQ KAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARE NQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDM YVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKK MKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQIL DSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAV VGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEIT LANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLG ITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGN ELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILA DANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKE VLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKKRKVGGGGSGAT NFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDA TYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYV QERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNV YIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSAL SKDPNEKRDHMVLLEFVTAAGITLGMDELYK BE4max w/o UGI, SEQ ID NO: 163 MKRTADGSEFESPKKKRKVSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCL LYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCG ECSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYC WRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIAL QSCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKY SIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATR LKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFG NIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNS DVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGL FGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKN LSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQ SKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIP HQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKS EETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTK VKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGV EDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLF DDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHD DSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKP ENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDN VPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETR QITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHH AHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYS NIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKT EVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKS KKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRM LASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEII EQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDT TIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKKR KVGGGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKF SVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDF FKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHK LEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPD NHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK nCas9 for BE4max-ΔUGI, SEQ ID NO: 164 MKRTADGSEFESPKKKRKVSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKK YSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEAT RLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIF GNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDN SDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNG LFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAK NLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFD QSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSI PHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRK SEETITPWNFEEWDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELT KVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISG VEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHL FDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIH DDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHK PENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSD NVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVET RQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYH HAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFY SNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKK TEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGK SKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKR MLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDE IIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFD TTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKK RKVGGGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHK FSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHD FFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGH KLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLP DNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK hA3A-BE3-ΔUGI-eUNG, SEQ ID NO: 165 MEASPASGPRHLMDPHIFTSNFNNGIGRHKTYLCYEVERLDNGTSVKMDQHRGFL HNQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISWSPCFSWGCAGEV RAFLQENTHVRLRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWDTFV DHQGCPFQPWDGLDEHSQALSGRLRAILQNQGNSGSETPGTSESATPESDKKYSI GLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLK RTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNI VDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNL SDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQS KNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSE ETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVE DRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFD DKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDD SLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLY YLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQI TKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIM NFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQ TGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKL KSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLAS AGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQI SEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTID RKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSANELTWHDVLAEEKQQP YFLNTLQTVASERQSGVTIYPPQKDVFNAFRFTELGDVKVVILGQDPYHGPGQAHG LAFSVRPGIAIPPSLLNMYKELENTIPGFTRPNHGYLESWARQGVLLLNTVLTVRAG QAHSHASLGWETFTDKVISLINQHREGVVFLLWGSHAQKKGAIIDKQRHHVLKAPH PSPLSAHRGFFGCNHFVLANQWLEQRGETPIDWMPVLPAESESGGSPKKKRKVG GGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVS GEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKS AMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEY NYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHY LSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK eA3A-BE3-ΔUGI-eUNG, SEQ ID NO: 166 MEASPASGPRHLMDPHIFTSNFNNGIGRHKTYLCYEVERLDNGTSVKMDQHRGFL HGQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISWSPCFSWGCAGEV RAFLQENTHVRLRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWDTFV DHQGCPFQPWDGLDEHSQALSGRLRAILQNQGNSGSETPGTSESATPESDKKYSI GLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLK RTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNI VDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNL SDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQS KNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSE ETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVE DRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFD DKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDD SLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLY YLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQI TKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIM NFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQ TGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKL KSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLAS AGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQI SEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTID RKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSANELTWHDVLAEEKQQP YFLNTLQTVASERQSGVTIYPPQKDVFNAFRFTELGDVKVVILGQDPYHGPGQAHG LAFSVRPGIAIPPSLLNMYKELENTIPGFTRPNHGYLESWARQGVLLLNTVLTVRAG QAHSHASLGWETFTDKVISLINQHREGVVFLLWGSHAQKKGAIIDKQRHHVLKAPH PSPLSAHRGFFGCNHFVLANQWLEQRGETPIDWMPVLPAESESGGSPKKKRKVG GGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVS GEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKS AMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEY NYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHY LSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK hAID-BE3-ΔUGI-eUNG, SEQ ID NO: 167 MDSLLMNRRKFLYQFKNVRWAKGRRETYLCYVVKRRDSATSFSLDFGYLRNKNGC HVELLFLRYISDWDLDPGRCYRVTWFTSWSPCYDCARHVADFLRGNPNLSLRIFTA RLYFCEDRKAEPEGLRRLHRAGVQIAIMTFKDYFYCWNTFVENHERTFKAWEGLHE NSVRLSRQLRRILLPLYEVDDLRDAFRTLGLSGSETPGTSESATPESDKKYSIGLAIG TNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTAR RRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEV AYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKL FIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIA LSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAIL LSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGY AGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLG ELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITP WNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVT EGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFN ASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKV MKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTF KEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIE MARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQN GRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEE VVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFK TEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGF SKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKE LLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQ KGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKR VILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTS TKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSANELTWHDVLAEEKQQPYFLNTL QTVASERQSGVTIYPPQKDVFNAFRFTELGDVKVVILGQDPYHGPGQAHGLAFSVR PGIAIPPSLLNMYKELENTIPGFTRPNHGYLESWARQGVLLLNTVLTVRAGQAHSHA SLGWETFTDKVISLINQHREGVVFLLWGSHAQKKGAIIDKQRHHVLKAPHPSPLSAH RGFFGCNHFVLANQWLEQRGETPIDWMPVLPAESESGGSPKKKRKVGGGGSGAT NFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDA TYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYV QERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNV YIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSAL SKDPNEKRDHMVLLEFVTAAGITLGMDELYK nCas9-eUNG for BE3, SEQ ID NO: 168 MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGET AEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHER HPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDL NPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGE KKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADL FLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYK EIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFD NGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAW MTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVY NELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSV EISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKT YAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMG RHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNE KLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGK SDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLV ETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINN YHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYF FYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIV KKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEK GKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGR KRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYL DEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKY FDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSANELTWHDVLAE EKQQPYFLNTLQTVASERQSGVTIYPPQKDVFNAFRFTELGDVKVVILGQDPYHGP GQAHGLAFSVRPGIAIPPSLLNMYKELENTIPGFTRPNHGYLESWARQGVLLLNTVL TVRAGQAHSHASLGWETFTDKVISLINQHREGVVFLLWGSHAQKKGAIIDKQRHHV LKAPHPSPLSAHRGFFGCNHFVLANQWLEQRGETPIDWMPVLPAESESGGSPKKK RKVGGGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHK FSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHD FFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGH KLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLP DNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK Target-AID-ΔUGI-eUNG, SEQ ID NO: 169 MAPKKKRKVGIHGVPAAMDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDR HSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFF HRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLA LAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSAR LSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDD DLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDL TLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELL VKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIP YYVGPLARGNSRFAWMTRKSEETITPWNFEEWDKGASAQSFIERMTNFDKNLPN EKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTV KQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIV LTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGK TILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKK GILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELG SQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKD DSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAER GGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLV SDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRK MIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRD FATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFD SPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKD LIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPED NEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENII HLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDS RADPKKKRKVGGGGTGGGGSAEYVRALFDFNGNDEEDLPFKKGDILRIRDKPEEQ WWNAEDSEGKRGMILVPYVEKYSGDYKDHDGDYKDHDIDYKDDDDKSGMTDAEY VRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGERRACFWGYAVNKPQSGTE RGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKILEWYNQELRGNGHTL KIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIFIQSSHNQLNENR WLEKTLKRAEKWRSELSIMIQVKILHTTKSPAVGPKKKRKVGTANELTWHDVLAEEK QQPYFLNTLQTVASERQSGVTIYPPQKDVFNAFRFTELGDVKVVILGQDPYHGPGQ AHGLAFSVRPGIAIPPSLLNMYKELENTIPGFTRPNHGYLESWARQGVLLLNTVLTV RAGQAHSHASLGWETFTDKVISLINQHREGVVFLLWGSHAQKKGAIIDKQRHHVLK APHPSPLSAHRGFFGCNHFVLANQWLEQRGETPIDWMPVLPAESEGGGGSGATN FSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDAT YGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQ ERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYI MADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSK DPNEKRDHMVLLEFVTAAGITLGMDELYK nCas9-eUNG for Target-AID, SEQ ID NO: 170 MAPKKKRKVGIHGVPAAMDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDR HSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFF HRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLA LAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSAR LSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDD DLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDL TLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELL VKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIP YYVGPLARGNSRFAWMTRKSEETITPWNFEEWDKGASAQSFIERMTNFDKNLPN EKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTV KQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIV LTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGK TILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKK GILQTVKWDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELG SQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKD DSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAER GGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLV SDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRK MIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRD FATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFD SPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKD LIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPED NEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENII HLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDS RADPKKKRKVGGGGTGGGGSAEYVRALFDFNGNDEEDLPFKKGDILRIRDKPEEQ WWNAEDSEGKRGMILVPYVEKYSGDYKDHDGDYKDHDIDYKDDDDKSGGPKKKR KVGTANELTWHDVLAEEKQQPYFLNTLQTVASERQSGVTIYPPQKDVFNAFRFTEL GDVKVVILGQDPYHGPGQAHGLAFSVRPGIAIPPSLLNMYKELENTIPGFTRPNHGY LESWARQGVLLLNTVLTVRAGQAHSHASLGWETFTDKVISLINQHREGVVFLLWGS HAQKKGAIIDKQRHHVLKAPHPSPLSAHRGFFGCNHFVLANQWLEQRGETPIDWM PVLPAESEGGGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDV NGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHM KQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDG NILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDG PVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK hA3A-BE3-ΔUGI-hUNG, SEQ ID NO: 171 MEASPASGPRHLMDPHIFTSNFNNGIGRHKTYLCYEVERLDNGTSVKMDQHRGFL HNQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISWSPCFSWGCAGEV RAFLQENTHVRLRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWDTFV DHQGCPFQPWDGLDEHSQALSGRLRAILQNQGNSGSETPGTSESATPESDKKYSI GLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLK RTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNI VDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNL SDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQS KNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSE ETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVE DRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFD DKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDD SLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLY YLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQI TKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIM NFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQ TGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKL KSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLAS AGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQI SEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTID RKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSIGQKTLYSFFSPSPARKR HAPSPEPAVQGTGVAGVPEESGDAAAIPAKKAPAGQEEPGTPPSSPLSAEQLDRIQ RNKAAALLRLAARNVPVGFGESWKKHLSGEFGKPYFIKLMGFVAEERKHYTVYPPP HQVFTWTQMCDIKDVKWILGQDPYHGPNQAHGLCFSVQRPVPPPPSLENIYKELS TDIEDFVHPGHGDLSGWAKQGVLLLNAVLTVRAHQANSHKERGWEQFTDAVVSWL NQNSNGLVFLLWGSYAQKKGSAIDRKRHHVLQTAHPSPLSVYRGFFGCRHFSKTN ELLQKSGKKPIDWKELSGGSPKKKRKVGGGGSGATNFSLLKQAGDVEENPGPMVS KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPT LVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFE GDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIED GSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGI TLGMDELYK eA3A-BE3-ΔUGI-hUNG, SEQ ID NO: 172 MEASPASGPRHLMDPHIFTSNFNNGIGRHKTYLCYEVERLDNGTSVKMDQHRGFL HGQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISWSPCFSWGCAGEV RAFLQENTHVRLRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWDTFV DHQGCPFQPWDGLDEHSQALSGRLRAILQNQGNSGSETPGTSESATPESDKKYSI GLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLK RTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNI VDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNL SDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQS KNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSE ETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVE DRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFD DKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDD SLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLY YLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQI TKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIM NFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQ TGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLWAKVEKGKSKKL KSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLAS AGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQI SEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTID RKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSIGQKTLYSFFSPSPARKR HAPSPEPAVQGTGVAGVPEESGDAAAIPAKKAPAGQEEPGTPPSSPLSAEQLDRIQ RNKAAALLRLAARNVPVGFGESWKKHLSGEFGKPYFIKLMGFVAEERKHYTVYPPP HQVFTWTQMCDIKDVKVVILGQDPYHGPNQAHGLCFSVQRPVPPPPSLENIYKELS TDIEDFVHPGHGDLSGWAKQGVLLLNAVLTVRAHQANSHKERGWEQFTDAVVSWL NQNSNGLVFLLWGSYAQKKGSAIDRKRHHVLQTAHPSPLSVYRGFFGCRHFSKTN ELLQKSGKKPIDWKELSGGSPKKKRKVGGGGSGATNFSLLKQAGDVEENPGPMVS KGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPT LVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFE GDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIED GSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGI TLGMDELYK hAID-BE3-ΔUGI-hUNG, SEQ ID NO: 173 MDSLLMNRRKFLYQFKNVRWAKGRRETYLCYVVKRRDSATSFSLDFGYLRNKNGC HVELLFLRYISDWDLDPGRCYRVTWFTSWSPCYDCARHVADFLRGNPNLSLRIFTA RLYFCEDRKAEPEGLRRLHRAGVQIAIMTFKDYFYCWNTFVENHERTFKAWEGLHE NSVRLSRQLRRILLPLYEVDDLRDAFRTLGLSGSETPGTSESATPESDKKYSIGLAIG TNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTAR RRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEV AYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKL FIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIA LSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAIL LSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGY AGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLG ELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITP WNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVT EGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFN ASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKV MKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTF KEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIE MARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQN GRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEE VVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFK TEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGF SKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKE LLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQ KGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKR VILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTS TKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSIGQKTLYSFFSPSPARKRHAPSPE PAVQGTGVAGVPEESGDAAAIPAKKAPAGQEEPGTPPSSPLSAEQLDRIQRNKAAA LLRLAARNVPVGFGESWKKHLSGEFGKPYFIKLMGFVAEERKHYTVYPPPHQVFT WTQMCDIKDVKVVILGQDPYHGPNQAHGLCFSVQRPVPPPPSLENIYKELSTDIEDF VHPGHGDLSGWAKQGVLLLNAVLTVRAHQANSHKERGWEQFTDAVVSWLNQNSN GLVFLLWGSYAQKKGSAIDRKRHHVLQTAHPSPLSVYRGFFGCRHFSKTNELLQKS GKKPIDWKELSGGSPKKKRKVGGGGSGATNFSLLKQAGDVEENPGPMVSKGEELF TGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLT YGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLV NRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQL ADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMD ELYK nCas9-hUNG for BE3, SEQ ID NO: 174 MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGET AEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHER HPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDL NPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGE KKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADL FLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYK EIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFD NGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAW MTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVY NELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSV EISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKT YAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMG RHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNE KLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGK SDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLV ETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINN YHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYF FYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIV KKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEK GKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGR KRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYL DEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKY FDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSIGQKTLYSFFSPS PARKRHAPSPEPAVQGTGVAGVPEESGDAAAIPAKKAPAGQEEPGTPPSSPLSAE QLDRIQRNKAAALLRLAARNVPVGFGESWKKHLSGEFGKPYFIKLMGFVAEERKHY TVYPPPHQVFTWTQMCDIKDVKVVILGQDPYHGPNQAHGLCFSVQRPVPPPPSLE NIYKELSTDIEDFVHPGHGDLSGWAKQGVLLLNAVLTVRAHQANSHKERGWEQFT DAVVSWLNQNSNGLVFLLWGSYAQKKGSAIDRKRHHVLQTAHPSPLSVYRGFFGC RHFSKTNELLQKSGKKPIDWKELSGGSPKKKRKVGGGGSGATNFSLLKQAGDVEE NPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTG KLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNY KTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVN FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK Target-AID-ΔUGI-hUNG, SEQ ID NO: 175 MAPKKKRKVGIHGVPAAMDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDR HSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFF HRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLA LAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSAR LSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDD DLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDL TLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELL VKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIP YYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPN EKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTV KQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIV LTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGK TILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKK GILQTVKWDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELG SQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKD DSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAER GGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLV SDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRK MIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRD FATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFD SPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKD LIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPED NEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENII HLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDS RADPKKKRKVGGGGTGGGGSAEYVRALFDFNGNDEEDLPFKKGDILRIRDKPEEQ WWNAEDSEGKRGMILVPYVEKYSGDYKDHDGDYKDHDIDYKDDDDKSGMTDAEY VRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGERRACFWGYAVNKPQSGTE RGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKILEWYNQELRGNGHTL KIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIFIQSSHNQLNENR WLEKTLKRAEKWRSELSIMIQVKILHTTKSPAVGPKKKRKVGTIGQKTLYSFFSPSPA RKRHAPSPEPAVQGTGVAGVPEESGDAAAIPAKKAPAGQEEPGTPPSSPLSAEQL DRIQRNKAAALLRLAARNVPVGFGESWKKHLSGEFGKPYFIKLMGFVAEERKHYTV YPPPHQVFTWTQMCDIKDVKVVILGQDPYHGPNQAHGLCFSVQRPVPPPPSLENIY KELSTDIEDFVHPGHGDLSGWAKQGVLLLNAVLTVRAHQANSHKERGWEQFTDAV VSWLNQNSNGLVFLLWGSYAQKKGSAIDRKRHHVLQTAHPSPLSVYRGFFGCRHF SKTNELLQKSGKKPIDWKELGGGGSGATNFSLLKQAGDVEENPGPMVSKGEELFT GVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTY GVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVN RIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLA DHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDE LYK nCas9-hUNG for Target-AID, SEQ ID NO: 176 MAPKKKRKVGIHGVPAAMDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDR HSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFF HRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLA LAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSAR LSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDD DLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDL TLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELL VKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIP YYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPN EKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTV KQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIV LTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGK TILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKK GILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELG SQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKD DSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAER GGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLV SDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRK MIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRD FATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFD SPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKD LIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPED NEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENII HLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDS RADPKKKRKVGGGGTGGGGSAEYVRALFDFNGNDEEDLPFKKGDILRIRDKPEEQ WWNAEDSEGKRGMILVPYVEKYSGDYKDHDGDYKDHDIDYKDDDDKSGGPKKKR KVGTIGQKTLYSFFSPSPARKRHAPSPEPAVQGTGVAGVPEESGDAAAIPAKKAPA GQEEPGTPPSSPLSAEQLDRIQRNKAAALLRLAARNVPVGFGESWKKHLSGEFGK PYFIKLMGFVAEERKHYTVYPPPHQVFTWTQMCDIKDVKVVILGQDPYHGPNQAHG LCFSVQRPVPPPPSLENIYKELSTDIEDFVHPGHGDLSGWAKQGVLLLNAVLTVRAH QANSHKERGWEQFTDAVVSWLNQNSNGLVFLLWGSYAQKKGSAIDRKRHHVLQT AHPSPLSVYRGFFGCRHFSKTNELLQKSGKKPIDWKELGGGGSGATNFSLLKQAG DVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFI CTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKD DGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKN GIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRD HMVLLEFVTAAGITLGMDELYK hA3A-BE3 w/o UGI, SEQ ID NO: 177 MEASPASGPRHLMDPHIFTSNFNNGIGRHKTYLCYEVERLDNGTSVKMDQHRGFL HNQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISWSPCFSWGCAGEV RAFLQENTHVRLRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWDTFV DHQGCPFQPWDGLDEHSQALSGRLRAILQNQGNSGSETPGTSESATPESDKKYSI GLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLK RTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNI VDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNL SDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQS KNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSE ETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVE DRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFD DKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDD SLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLY YLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQI TKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIM NFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQ TGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLWAKVEKGKSKKL KSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLAS AGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQI SEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTID RKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSPKKKRKVGGGGSGATNF SLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATY GKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQE RTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIM ADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKD PNEKRDHMVLLEFVTAAGITLGMDELYK eA3A-BE3 w/o UGI, SEQ ID NO: 178 MEASPASGPRHLMDPHIFTSNFNNGIGRHKTYLCYEVERLDNGTSVKMDQHRGFL HGQAKNLLCGFYGRHAELRFLDLVPSLQLDPAQIYRVTWFISWSPCFSWGCAGEV RAFLQENTHVRLRIFAARIYDYDPLYKEALQMLRDAGAQVSIMTYDEFKHCWDTFV DHQGCPFQPWDGLDEHSQALSGRLRAILQNQGNSGSETPGTSESATPESDKKYSI GLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLK RTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNI VDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNL SDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQS KNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSE ETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVE DRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFD DKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDD SLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLY YLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQI TKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIM NFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQ TGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKL KSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLAS AGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQI SEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTID RKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSPKKKRKVGGGGSGATNF SLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATY GKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQE RTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIM ADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKD PNEKRDHMVLLEFVTAAGITLGMDELYK hAID-BE3 w/o UGI, SEQ ID NO: 179 MDSLLMNRRKFLYQFKNVRWAKGRRETYLCYVVKRRDSATSFSLDFGYLRNKNGC HVELLFLRYISDWDLDPGRCYRVTWFTSWSPCYDCARHVADFLRGNPNLSLRIFTA RLYFCEDRKAEPEGLRRLHRAGVQIAIMTFKDYFYCWNTFVENHERTFKAWEGLHE NSVRLSRQLRRILLPLYEVDDLRDAFRTLGLSGSETPGTSESATPESDKKYSIGLAIG TNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTAR RRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEV AYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKL FIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIA LSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAIL LSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGY AGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLG ELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITP WNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVT EGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFN ASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKV MKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTF KEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIE MARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQN GRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEE VVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHV AQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYL NAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFK TEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGF SKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKE LLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQ KGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKR VILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTS TKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSPKKKRKVGGGGSGATNFSLLKQA GDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKD DGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKN GIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRD HMVLLEFVTAAGITLGMDELYK nCas9 for BE3 w/o UGI, SEQ ID NO: 180 MDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGET AEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHER HPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDL NPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGE KKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADL FLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYK EIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFD NGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAW MTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVY NELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSV EISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKT YAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMG RHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNE KLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGK SDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLV ETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINN YHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYF FYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIV KKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEK GKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGR KRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYL DEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKY FDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSPKKKRKVGGGG SGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEG EGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMP EGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYN SHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLST QSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK Target-AID w/o UGI, SEQ ID NO: 181 MAPKKKRKVGIHGVPAAMDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDR HSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFF HRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLA LAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSAR LSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDD DLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDL TLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELL VKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIP YYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPN EKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTV KQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIV LTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGK TILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKK GILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELG SQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKD DSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAER GGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLV SDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRK MIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRD FATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFD SPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKD LIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPED NEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENII HLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDS RADPKKKRKVGGGGTGGGGSAEYVRALFDFNGNDEEDLPFKKGDILRIRDKPEEQ WWNAEDSEGKRGMILVPYVEKYSGDYKDHDGDYKDHDIDYKDDDDKSGMTDAEY VRIHEKLDIYTFKKQFFNNKKSVSHRCYVLFELKRRGERRACFWGYAVNKPQSGTE RGIHAEIFSIRKVEEYLRDNPGQFTINWYSSWSPCADCAEKILEWYNQELRGNGHTL KIWACKLYYEKNARNQIGLWNLRDNGVGLNVMVSEHYQCCRKIFIQSSHNQLNENR WLEKTLKRAEKWRSELSIMIQVKILHTTKSPAVGPKKKRKVGGGGSGATNFSLLKQ AGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFF KDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQ KNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEK RDHMVLLEFVTAAGITLGMDELYK nCas9 for Target-AID w/o UGI, SEQ ID NO: 182 MAPKKKRKVGIHGVPAAMDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDR HSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFF HRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLA LAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSAR LSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDD DLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDL TLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELL VKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIP YYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPN EKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTV KQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIV LTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGK TILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKK GILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELG SQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKD DSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAER GGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLV SDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRK MIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRD FATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFD SPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKD LIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPED NEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENII HLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDS RADPKKKRKVGGGGTGGGGSAEYVRALFDFNGNDEEDLPFKKGDILRIRDKPEEQ WWNAEDSEGKRGMILVPYVEKYSGDYKDHDGDYKDHDIDYKDDDDKSGGPKKKR KVGGGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKF SVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDF FKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHK LEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPD NHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDE SPACE-ΔUGI-eUNG, SEQ ID NO: 183 MKRTADGSEFESPKKKRKVSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLN NRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYGTFEPCVMCAGA MIHSRIGRVVFGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALLCYFFR MPRQVFNAQKKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYS IGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRL KRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNL SDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQS KNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSE ETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVE DRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFD DKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDD SLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLY YLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQI TKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIM NFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQ TGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKL KSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLAS AGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQI SEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTID RKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKKRKV SGGSSGGSSGSETPGTSESATPESSGGSSGGSTDAEYVRIHEKLDIYTFKKQFFNN KKSVSHRCYVLFELKRRGERRACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRD NPGQFTINWYSSWSPCADCAEKILEWYNQELRGNGHTLKIWACKLYYEKNARNQI GLWNLRDNGVGLNVMVSEHYQCCRKIFIQSSHNQLNENRWLEKTLKRAEKWRSEL SIMIQVKILHTTKSPAVSGGSGGSGGSANELTWHDVLAEEKQQPYFLNTLQTVASE RQSGVTIYPPQKDVFNAFRFTELGDVKVVILGQDPYHGPGQAHGLAFSVRPGIAIPP SLLNMYKELENTIPGFTRPNHGYLESWARQGVLLLNTVLTVRAGQAHSHASLGWET FTDKVISLINQHREGVVFLLWGSHAQKKGAIIDKQRHHVLKAPHPSPLSAHRGFFGC NHFVLANQWLEQRGETPIDWMPVLPAESEGSGATNFSLLKQAGDVEENPGPMVSK GEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTL VTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFE GDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIED GSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGI TLGMDELYKSGGSPKKKRKV SPACE-ΔUGI-hUNG, SEQ ID NO: 184 MKRTADGSEFESPKKKRKVSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLN NRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYGTFEPCVMCAGA MIHSRIGRVVFGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALLCYFFR MPRQVFNAQKKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYS IGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRL KRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNL SDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQS KNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSE ETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVE DRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFD DKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDD SLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLY YLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQI TKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIM NFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQ TGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKL KSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLAS AGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQI SEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTID RKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKKRKV SGGSSGGSSGSETPGTSESATPESSGGSSGGSTDAEYVRIHEKLDIYTFKKQFFNN KKSVSHRCYVLFELKRRGERRACFWGYAVNKPQSGTERGIHAEIFSIRKVEEYLRD NPGQFTINWYSSWSPCADCAEKILEWYNQELRGNGHTLKIWACKLYYEKNARNQI GLWNLRDNGVGLNVMVSEHYQCCRKIFIQSSHNQLNENRWLEKTLKRAEKWRSEL SIMIQVKILHTTKSPAVSGGSGGSGGSIGQKTLYSFFSPSPARKRHAPSPEPAVQGT GVAGVPEESGDAAAIPAKKAPAGQEEPGTPPSSPLSAEQLDRIQRNKAAALLRLAA RNVPVGFGESWKKHLSGEFGKPYFIKLMGFVAEERKHYTVYPPPHQVFTWTQMCD IKDVKVVILGQDPYHGPNQAHGLCFSVQRPVPPPPSLENIYKELSTDIEDFVHPGHG DLSGWAKQGVLLLNAVLTVRAHQANSHKERGWEQFTDAVVSWLNQNSNGLVFLL WGSYAQKKGSAIDRKRHHVLQTAHPSPLSVYRGFFGCRHFSKTNELLQKSGKKPID WKELGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFS VSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFF KSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKL EYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDN HYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKSGGSPKKKRKV ABEmax-eUNG, SEQ ID NO: 185 MKRTADGSEFESPKKKRKVSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVH NNRVIGEGWNRPIGRHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTLEPCVMCAG AMIHSRIGRVVFGARDAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLSDFF RMRRQEIKAQKKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSSEVE FSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIM ALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNAKTGAAGSL MDVLHYPGMNHRVEITEGILADECAALLCYFFRMPRQVFNAQKKAQSSTDSGGSS GGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYKVPSK KFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSN EMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDS TDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINAS GVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDA KLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASM IKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPI LEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDN REKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIER MTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDL LFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDN EENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLI NGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHI ANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENMEMARENQTTQKGQKNSRER MKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYD VDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLIT QRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLI REVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEF VYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNG ETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKK DWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDF LEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLA SHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKH RDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYE TRIDLSQLGGDSGGSGGSGGSANELTWHDVLAEEKQQPYFLNTLQTVASERQSGV TIYPPQKDVFNAFRFTELGDVKVVILGQDPYHGPGQAHGLAFSVRPGIAIPPSLLNM YKELENTIPGFTRPNHGYLESWARQGVLLLNTVLTVRAGQAHSHASLGWETFTDKV ISLINQHREGVVFLLWGSHAQKKGAIIDKQRHHVLKAPHPSPLSAHRGFFGCNHFVL ANQWLEQRGETPIDWMPVLPAESESGGSKRTADGSEFEPKKKRKVGSGATNFSLL KQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTI FFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMAD KQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPN EKRDHMVLLEFVTAAGITLGMDELYKSGGSPKKKRKV miniABEmax(V82G)-eUNG, SEQ ID NO: 186 MKRTADGSEFESPKKKRKVSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLN NRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYGTFEPCVMCAGA MIHSRIGRWFGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALLCYFFR MPRQVFNAQKKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYS IGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRL KRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNL SDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQS KNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSE ETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVE DRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFD DKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDD SLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLY YLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQI TKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIM NFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQ TGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKL KSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLAS AGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQI SEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTID RKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSGGSGGSANELTWHDVL AEEKQQPYFLNTLQTVASERQSGVTIYPPQKDVFNAFRFTELGDVKVVILGQDPYH GPGQAHGLAFSVRPGIAIPPSLLNMYKELENTIPGFTRPNHGYLESWARQGVLLLNT VLTVRAGQAHSHASLGWETFTDKVISLINQHREGVVFLLWGSHAQKKGAIIDKQRH HVLKAPHPSPLSAHRGFFGCNHFVLANQWLEQRGETPIDWMPVLPAESESGGSKR TADGSEFEPKKKRKVGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVEL DGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRY PDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDF KEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTP IGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKSGGSP KKKRKV nCas9-eUNG for ABEmax, SEQ ID NO: 187 MKRTADGSEFESPKKKRKVSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKK YSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEAT RLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIF GNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDN SDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNG LFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAK NLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFD QSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSI PHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRK SEETITPWNFEEWDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELT KVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISG VEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHL FDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIH DDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHK PENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSD NVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVET RQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYH HAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFY SNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKK TEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGK SKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKR MLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDE IIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFD TTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSGGSGGSANELTWH DVLAEEKQQPYFLNTLQTVASERQSGVTIYPPQKDVFNAFRFTELGDVKVVILGQDP YHGPGQAHGLAFSVRPGIAIPPSLLNMYKELENTIPGFTRPNHGYLESWARQGVLLL NTVLTVRAGQAHSHASLGWETFTDKVISLINQHREGVVFLLWGSHAQKKGAIIDKQR HHVLKAPHPSPLSAHRGFFGCNHFVLANQWLEQRGETPIDWMPVLPAESESGGSK RTADGSEFEPKKKRKVGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVE LDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSR YPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGID FKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNT PIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKSGGS PKKKRKV eUNG-ABEmax, SEQ ID NO: 188 MKRTADGSEFESPKKKRKVANELTWHDVLAEEKQQPYFLNTLQTVASERQSGVTIY PPQKDVFNAFRFTELGDVKVVILGQDPYHGPGQAHGLAFSVRPGIAIPPSLLNMYKE LENTIPGFTRPNHGYLESWARQGVLLLNTVLTVRAGQAHSHASLGWETFTDKVISLI NQHREGVVFLLWGSHAQKKGAIIDKQRHHVLKAPHPSPLSAHRGFFGCNHFVLAN QWLEQRGETPIDWMPVLPAESESGGSGGSGGSSEVEFSHEYWMRHALTLAKRA WDEREVPVGAVLVHNNRVIGEGWNRPIGRHDPTAHAEIMALRQGGLVMQNYRLID ATLYVTLEPCVMCAGAMIHSRIGRVVFGARDAKTGAAGSLMDVLHHPGMNHRVEIT EGILADECAALLSDFFRMRRQEIKAQKKAQSSTDSGGSSGGSSGSETPGTSESATP ESSGGSSGGSSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGW NRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRV VFGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALLCYFFRMPRQVFNA QKKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNS VGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRY TRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYH EKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQL VQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSL GLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSD ILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYI DGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHA ILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFE EVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMR KPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGT YHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLK RRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQ KAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARE NQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDM YVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKK MKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQIL DSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAV VGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEIT LANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLG ITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGN ELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILA DANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKE VLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKKRKVGSGATNFSL LKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGK LTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTI FFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMAD KQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPN EKRDHMVLLEFVTAAGITLGMDELYKSGGSPKKKRKV eUNG-miniABEmax(V82G), SEQ ID NO: 189 MKRTADGSEFESPKKKRKVANELTWHDVLAEEKQQPYFLNTLQTVASERQSGVTIY PPQKDVFNAFRFTELGDVKVVILGQDPYHGPGQAHGLAFSVRPGIAIPPSLLNMYKE LENTIPGFTRPNHGYLESWARQGVLLLNTVLTVRAGQAHSHASLGWETFTDKVISLI NQHREGVVFLLWGSHAQKKGAIIDKQRHHVLKAPHPSPLSAHRGFFGCNHFVLAN QWLEQRGETPIDWMPVLPAESESGGSGGSGGSSEVEFSHEYWMRHALTLAKRAR DEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDAT LYGTFEPCVMCAGAMIHSRIGRWFGVRNAKTGAAGSLMDVLHYPGMNHRVEITE GILADECAALLCYFFRMPRQVFNAQKKAQSSTDSGGSSGGSSGSETPGTSESATP ESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLI GALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLA QIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALV RQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNRE DLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPL ARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPK HSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKED YFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFE DREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLK SDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTV KWDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKE HPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDN KVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLS ELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRK DFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKS EQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATV RKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTV AYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKL PKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQ KQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFT LTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGS KRTADGSEFEPKKKRKVGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILV ELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFS RYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGI DFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQN TPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKSGGS PKKKRKV eUNG-nCas9 for ABEmax, SEQ ID NO: 190 MKRTADGSEFESPKKKRKVANELTWHDVLAEEKQQPYFLNTLQTVASERQSGVTIY PPQKDVFNAFRFTELGDVKVVILGQDPYHGPGQAHGLAFSVRPGIAIPPSLLNMYKE LENTIPGFTRPNHGYLESWARQGVLLLNTVLTVRAGQAHSHASLGWETFTDKVISLI NQHREGVVFLLWGSHAQKKGAIIDKQRHHVLKAPHPSPLSAHRGFFGCNHFVLAN QWLEQRGETPIDWMPVLPAESESGGSGGSGGSSGGSSGGSSGSETPGTSESATP ESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLI GALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESF LVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKF RGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRL ENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLA QIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALV RQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNRE DLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPL ARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPK HSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKED YFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFE DREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLK SDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTV KVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKE HPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDN KVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLS ELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRK DFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKS EQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATV RKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTV AYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKL PKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQ KQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFT LTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGS KRTADGSEFEPKKKRKVGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILV ELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFS RYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGI DFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQN TPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKSGGS PKKKRKV ABEmax-hUNG, SEQ ID NO: 191 MKRTADGSEFESPKKKRKVSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVH NNRVIGEGWNRPIGRHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTLEPCVMCAG AMIHSRIGRVVFGARDAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLSDFF RMRRQEIKAQKKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSSEVE FSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIM ALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNAKTGAAGSL MDVLHYPGMNHRVEITEGILADECAALLCYFFRMPRQVFNAQKKAQSSTDSGGSS GGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYKVPSK KFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSN EMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDS TDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINAS GVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDA KLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASM IKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPI LEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDN REKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIER MTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDL LFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDN EENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLI NGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHI ANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRER MKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYD VDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLIT QRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLI REVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEF VYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNG ETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKK DWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDF LEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLA SHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKH RDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYE TRIDLSQLGGDSGGSGGSGGSIGQKTLYSFFSPSPARKRHAPSPEPAVQGTGVAG VPEESGDAAAIPAKKAPAGQEEPGTPPSSPLSAEQLDRIQRNKAAALLRLAARNVPV GFGESWKKHLSGEFGKPYFIKLMGFVAEERKHYTVYPPPHQVFTWTQMCDIKDVK VVILGQDPYHGPNQAHGLCFSVQRPVPPPPSLENIYKELSTDIEDFVHPGHGDLSG WAKQGVLLLNAVLTVRAHQANSHKERGWEQFTDAVVSWLNQNSNGLVFLLWGSY AQKKGSAIDRKRHHVLQTAHPSPLSVYRGFFGCRHFSKTNELLQKSGKKPIDWKEL SGGSKRTADGSEFEPKKKRKVGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGV VPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGV QCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIE LKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHY QQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK SGGSPKKKRKV miniABEmax(V82G)-hUNG, SEQ ID NO: 192 MKRTADGSEFESPKKKRKVSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLN NRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYGTFEPCVMCAGA MIHSRIGRVVFGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALLCYFFR MPRQVFNAQKKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYS IGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRL KRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNL SDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQS KNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSE ETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVE DRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFD DKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDD SLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLY YLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQI TKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIM NFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQ TGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKL KSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLAS AGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQI SEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTID RKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSGGSGGSIGQKTLYSFFS PSPARKRHAPSPEPAVQGTGVAGVPEESGDAAAIPAKKAPAGQEEPGTPPSSPLS AEQLDRIQRNKAAALLRLAARNVPVGFGESWKKHLSGEFGKPYFIKLMGFVAEERK HYTVYPPPHQVFTWTQMCDIKDVKVVILGQDPYHGPNQAHGLCFSVQRPVPPPPS LENIYKELSTDIEDFVHPGHGDLSGWAKQGVLLLNAVLTVRAHQANSHKERGWEQF TDAVVSWLNQNSNGLVFLLWGSYAQKKGSAIDRKRHHVLQTAHPSPLSVYRGFFG CRHFSKTNELLQKSGKKPIDWKELSGGSKRTADGSEFEPKKKRKVGSGATNFSLLK QAGDVEENPGPMVSKGEELFTGWPILVELDGDVNGHKFSVSGEGEGDATYGKLT LKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIF FKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADK QKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNE KRDHMVLLEFVTAAGITLGMDELYKSGGSPKKKRKV nCas9-hUNG for ABEmax, SEQ ID NO: 193 MKRTADGSEFESPKKKRKVSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKK YSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEAT RLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIF GNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDN SDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNG LFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAK NLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFD QSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSI PHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRK SEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELT KVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISG VEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHL FDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIH DDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHK PENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSD NVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVET RQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYH HAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFY SNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKK TEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGK SKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKR MLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDE IIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFD TTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSGGSGGSIGQKTLYS FFSPSPARKRHAPSPEPAVQGTGVAGVPEESGDAAAIPAKKAPAGQEEPGTPPSS PLSAEQLDRIQRNKAAALLRLAARNVPVGFGESWKKHLSGEFGKPYFIKLMGFVAE ERKHYTVYPPPHQVFTWTQMCDIKDVKVVILGQDPYHGPNQAHGLCFSVQRPVPP PPSLENIYKELSTDIEDFVHPGHGDLSGWAKQGVLLLNAVLTVRAHQANSHKERGW EQFTDAVVSWLNQNSNGLVFLLWGSYAQKKGSAIDRKRHHVLQTAHPSPLSVYRG FFGCRHFSKTNELLQKSGKKPIDWKELSGGSKRTADGSEFEPKKKRKVGSGATNF SLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATY GKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQE RTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIM ADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKD PNEKRDHMVLLEFVTAAGITLGMDELYKSGGSPKKKRKV hUNG-ABEmax, SEQ ID NO: 194 MKRTADGSEFESPKKKRKVIGQKTLYSFFSPSPARKRHAPSPEPAVQGTGVAGVP EESGDAAAIPAKKAPAGQEEPGTPPSSPLSAEQLDRIQRNKAAALLRLAARNVPVG FGESWKKHLSGEFGKPYFIKLMGFVAEERKHYTVYPPPHQVFTWTQMCDIKDVKV VILGQDPYHGPNQAHGLCFSVQRPVPPPPSLENIYKELSTDIEDFVHPGHGDLSGW AKQGVLLLNAVLTVRAHQANSHKERGWEQFTDAVVSWLNQNSNGLVFLLWGSYA QKKGSAIDRKRHHVLQTAHPSPLSVYRGFFGCRHFSKTNELLQKSGKKPIDWKELS GGSGGSGGSSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVHNNRVIGEGW NRPIGRHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTLEPCVMCAGAMIHSRIGRV VFGARDAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLSDFFRMRRQEIKA QKKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSSEVEFSHEYWMRH ALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVM QNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNAKTGAAGSLMDVLHYPGM NHRVEITEGILADECAALLCYFFRMPRQVFNAQKKAQSSTDSGGSSGGSSGSETPG TSESATPESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDR HSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFF HRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLA LAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSAR LSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDD DLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDL TLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELL VKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIP YYVGPLARGNSRFAWMTRKSEETITPWNFEEWDKGASAQSFIERMTNFDKNLPN EKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTV KQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIV LTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGK TILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKK GILQTVKWDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELG SQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKD DSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAER GGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLV SDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRK MIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRD FATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFD SPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKD LIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPED NEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENII HLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDS GGSKRTADGSEFEPKKKRKVGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVV PILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQ CFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIEL KGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHY QQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK SGGSPKKKRKV hUNG-miniABEmax(V82G), SEQ ID NO: 195 MKRTADGSEFESPKKKRKVIGQKTLYSFFSPSPARKRHAPSPEPAVQGTGVAGVP EESGDAAAIPAKKAPAGQEEPGTPPSSPLSAEQLDRIQRNKAAALLRLAARNVPVG FGESWKKHLSGEFGKPYFIKLMGFVAEERKHYTVYPPPHQVFTWTQMCDIKDVKV VILGQDPYHGPNQAHGLCFSVQRPVPPPPSLENIYKELSTDIEDFVHPGHGDLSGW AKQGVLLLNAVLTVRAHQANSHKERGWEQFTDAVVSWLNQNSNGLVFLLWGSYA QKKGSAIDRKRHHVLQTAHPSPLSVYRGFFGCRHFSKTNELLQKSGKKPIDWKELS GGSGGSGGSSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWN RAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYGTFEPCVMCAGAMIHSRIGRVV FGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALLCYFFRMPRQVFNAQ KKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNSV GWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYT RRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHE KYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLV QTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLG LTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDIL RVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYID GGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAI LRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFE EVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMR KPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGT YHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLK RRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQ KAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARE NQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDM YVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKK MKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQIL DSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAV VGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEIT LANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLG ITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGN ELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILA DANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKE VLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKKRKVGSGATNFSL LKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGK LTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTI FFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMAD KQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPN EKRDHMVLLEFVTAAGITLGMDELYKSGGSPKKKRKV hUNG-nCas9 for ABEmax, SEQ ID NO: 196 MKRTADGSEFESPKKKRKVIGQKTLYSFFSPSPARKRHAPSPEPAVQGTGVAGVP EESGDAAAIPAKKAPAGQEEPGTPPSSPLSAEQLDRIQRNKAAALLRLAARNVPVG FGESWKKHLSGEFGKPYFIKLMGFVAEERKHYTVYPPPHQVFTWTQMCDIKDVKV VILGQDPYHGPNQAHGLCFSVQRPVPPPPSLENIYKELSTDIEDFVHPGHGDLSGW AKQGVLLLNAVLTVRAHQANSHKERGWEQFTDAVVSWLNQNSNGLVFLLWGSYA QKKGSAIDRKRHHVLQTAHPSPLSVYRGFFGCRHFSKTNELLQKSGKKPIDWKELS GGSGGSGGSSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNS VGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRY TRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYH EKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQL VQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSL GLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSD ILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYI DGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHA ILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFE EVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMR KPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGT YHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLK RRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQ KAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENIVIEMARE NQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDM YVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKK MKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQIL DSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAV VGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEIT LANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKE SILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLG ITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGN ELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILA DANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKE VLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKKRKVGSGATNFSL LKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGK LTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTI FFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMAD KQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPN EKRDHMVLLEFVTAAGITLGMDELYKSGGSPKKKRKV ABEmax-UGI, SEQ ID NO: 197 MKRTADGSEFESPKKKRKVSEVEFSHEYWMRHALTLAKRAWDEREVPVGAVLVH NNRVIGEGWNRPIGRHDPTAHAEIMALRQGGLVMQNYRLIDATLYVTLEPCVMCAG AMIHSRIGRVVFGARDAKTGAAGSLMDVLHHPGMNHRVEITEGILADECAALLSDFF RMRRQEIKAQKKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSSEVE FSHEYWMRHALTLAKRARDEREVPVGAVLVLNNRVIGEGWNRAIGLHDPTAHAEIM ALRQGGLVMQNYRLIDATLYVTFEPCVMCAGAMIHSRIGRVVFGVRNAKTGAAGSL MDVLHYPGMNHRVEITEGILADECAALLCYFFRMPRQVFNAQKKAQSSTDSGGSS GGSSGSETPGTSESATPESSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYKVPSK KFKVLGNTDRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSN EMAKVDDSFFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDS TDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINAS GVDAKAILSARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDA KLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASM IKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPI LEKMDGTEELLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDN REKIEKILTFRIPYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIER MTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDL LFKTNRKVTVKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDN EENEDILEDIVLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLI NGIRDKQSGKTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHI ANLAGSPAIKKGILQTVKVVDELVKVMGRHKPENMEMARENQTTQKGQKNSRER MKRIEEGIKELGSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYD VDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLIT QRKFDNLTKAERGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLI REVKVITLKSKLVSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEF VYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNG ETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKK DWDPKKYGGFDSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDF LEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLA SHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKH RDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYE TRIDLSQLGGDSGGSGGSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPE SDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLSGGSGGSGGS TNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTS DAPEYKPWALVIQDSNGENKIKMLSGGSKRTADGSEFEPKKKRKVGSGATNFSLLK QAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLT LKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIF FKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADK QKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNE KRDHMVLLEFVTAAGITLGMDELYKSGGSPKKKRKV miniABEmax(V82G)-UGI, SEQ ID NO: 198 MKRTADGSEFESPKKKRKVSEVEFSHEYWMRHALTLAKRARDEREVPVGAVLVLN NRVIGEGWNRAIGLHDPTAHAEIMALRQGGLVMQNYRLIDATLYGTFEPCVMCAGA MIHSRIGRVVFGVRNAKTGAAGSLMDVLHYPGMNHRVEITEGILADECAALLCYFFR MPRQVFNAQKKAQSSTDSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYS IGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRL KRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNL SDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQS KNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSE ETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVE DRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFD DKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDD SLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLY YLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQI TKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIM NFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQ TGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLWAKVEKGKSKKL KSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLAS AGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQI SEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTID RKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSGGSGGSTNLSDIIEKETG KQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALV IQDSNGENKIKMLSGGSGGSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVIGNK PESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLSGGSKRTAD GSEFEPKKKRKVGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGD VNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDH MKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKED GNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGD GPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKSGGSPKKK RKV nCas9-UGI for ABEmax, SEQ ID NO: 199 MKRTADGSEFESPKKKRKVSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKK YSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEAT RLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIF GNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDN SDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNG LFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAK NLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFD QSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSI PHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRK SEETITPWNFEEWDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELT KVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISG VEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHL FDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIH DDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHK PENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSD NVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVET RQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYH HAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFY SNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKK TEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGK SKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKR MLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDE IIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFD TTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSGGSGGSTNLSDIIEK ETGKQLVIQESILMLPEEVEEVIGNKPESDILVHTAYDESTDENVMLLTSDAPEYKPW ALVIQDSNGENKIKMLSGGSGGSGGSTNLSDIIEKETGKQLVIQESILMLPEEVEEVI GNKPESDILVHTAYDESTDENVMLLTSDAPEYKPWALVIQDSNGENKIKMLSGGSK RTADGSEFEPKKKRKVGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVE LDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSR YPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGID FKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNT PIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKSGGS PKKKRKV REV1 (human) amino acid sequence, SEQ ID NO: 200 MRRGGWRKRAENDGWETWGGYMAAKVQKLEEQFRSDAAMQKDGTSSTIFSGVA IYVNGYTDPSAEELRKLMMLHGGQYHVYYSRSKTTHIIATNLPNAKIKELKGEKVIRP EWIVESIKAGRLLSYIPYQLYTKQSSVQKGLSFNPVCRPEDPLPGPSNIAKQLNNRV NHIVKKIETENEVKVNGMNSWNEEDENNDFSFVDLEQTSPGRKQNGIPHPRGSTAI FNGHTPSSNGALKTQDCLVPMVNSVASRLSPAFSQEEDKAEKSSTDFRDCTLQQL QQSTRNTDALRNPHRTNSFSLSPLHSNTKINGAHHSTVQGPSSTKSTSSVSTFSKA APSVPSKPSDCNFISNFYSHSRLHHISMWKCELTEFVNTLQRQSNGIFPGREKLKK MKTGRSALVVTDTGDMSVLNSPRHQSCIMHVDMDCFFVSVGIRNRPDLKGKPVAV TSNRGTGRAPLRPGANPQLEWQYYQNKILKGKAADIPDSSLWENPDSAQANGIDS VLSRAEIASCSYEARQLGIKNGMFFGHAKQLCPNLQAVPYDFHAYKEVAQTLYETLA SYTHNIEAVSCDEALVDITEILAETKLTPDEFANAVRMEIKDQTKCAASVGIGSNILLA RMATRKAKPDGQYHLKPEEVDDFIRGQLVTNLPGVGHSMESKLASLGIKTCGDLQY MTMAKLQKEFGPKTGQMLYRFCRGLDDRPVRTEKERKSVSAEINYGIRFTQPKEAE AFLLSLSEEIQRRLEATGMKGKRLTLKIMVRKPGAPVETAKFGGHGICDNIARTVTLD QATDNAKIIGKAMLNMFHTMKLNISDMRGVGIHVNQLVPTNLNPSTCPSRPSVQSS HFPSGSYSVRDVFQVQKAKKSTEEEHKEVFRAAVDLEISSASRTCTFLPPFPAHLPT SPDTNKAESSGKWNGLHTPVSVQSRLNLSIEVPSPSQLDQSVLEALPPDLREQVEQ VCAVQQAESHGDKKKEPVNGCNTGILPQPVGTVLLQIPEPQESNSDAGINLIALPAF SQVDPEVFAALPAELQRELKAAYDQRQRQGENSTHQQSASASVPKNPLLHLKAAV KEKKRNKKKKTIGSPKRIQSPLNNKLLNSPAKTLPGACGSPQKLIDGFLKHEGPPAE KPLEELSASTSGVPGLSSLQSDPAGCVRPPAPNLAGAVEFNDVKTLLREWITTISDP MEEDILQVVKYCTDLIEEKDLEKLDLVIKYMKRLMQQSVESVWNMAFDFILDNVQVV LQQTYGSTLKVT BE4max-REV1, SEQ ID NO: 201 MKRTADGSEFESPKKKRKVSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCL LYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCG ECSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYC WRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIAL QSCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKY SIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATR LKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFG NIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNS DVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGL FGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKN LSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQ SKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIP HQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKS EETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTK VKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGV EDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLF DDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHD DSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKP ENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDN VPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETR QITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHH AHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYS NIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKT EVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKS KKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRM LASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEII EQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDT TIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSRRGGWRKRAENDG WETWGGYMAAKVQKLEEQFRSDAAMQKDGTSSTIFSGVAIYVNGYTDPSAEELRK LMMLHGGQYHVYYSRSKTTHIIATNLPNAKIKELKGEKVIRPEWIVESIKAGRLLSYIP YQLYTKQSSVQKGLSFNPVCRPEDPLPGPSNIAKQLNNRVNHIVKKIETENEVKVNG MNSWNEEDENNDFSFVDLEQTSPGRKQNGIPHPRGSTAIFNGHTPSSNGALKTQD CLVPMVNSVASRLSPAFSQEEDKAEKSSTDFRDCTLQQLQQSTRNTDALRNPHRT NSFSLSPLHSNTKINGAHHSTVQGPSSTKSTSSVSTFSKAAPSVPSKPSDCNFISNF YSHSRLHHISMWKCELTEFVNTLQRQSNGIFPGREKLKKMKTGRSALVVTDTGDMS VLNSPRHQSCIMHVDMDCFFVSVGIRNRPDLKGKPVAVTSNRGTGRAPLRPGANP QLEWQYYQNKILKGKAADIPDSSLWENPDSAQANGIDSVLSRAEIASCSYEARQLGI KNGMFFGHAKQLCPNLQAVPYDFHAYKEVAQTLYETLASYTHNIEAVSCDEALVDIT EILAETKLTPDEFANAVRMEIKDQTKCAASVGIGSNILLARMATRKAKPDGQYHLKPE EVDDFIRGQLVTNLPGVGHSMESKLASLGIKTCGDLQYMTMAKLQKEFGPKTGQML YRFCRGLDDRPVRTEKERKSVSAEINYGIRFTQPKEAEAFLLSLSEEIQRRLEATGM KGKRLTLKIMVRKPGAPVETAKFGGHGICDNIARTVTLDQATDNAKIIGKAMLNMFH TMKLNISDMRGVGIHVNQLVPTNLNPSTCPSRPSVQSSHFPSGSYSVRDVFQVQKA KKSTEEEHKEVFRAAVDLEISSASRTCTFLPPFPAHLPTSPDTNKAESSGKWNGLHT PVSVQSRLNLSIEVPSPSQLDQSVLEALPPDLREQVEQVCAVQQAESHGDKKKEPV NGCNTGILPQPVGTVLLQIPEPQESNSDAGINLIALPAFSQVDPEVFAALPAELQREL KAAYDQRQRQGENSTHQQSASASVPKNPLLHLKAAVKEKKRNKKKKTIGSPKRIQS PLNNKLLNSPAKTLPGACGSPQKLIDGFLKHEGPPAEKPLEELSASTSGVPGLSSLQ SDPAGCVRPPAPNLAGAVEFNDVKTLLREWITTISDPMEEDILQVVKYCTDLIEEKDL EKLDLVIKYMKRLMQQSVESVWNMAFDFILDNVQVVLQQTYGSTLKVTSGGSKRTA DGSEFEPKKKRKVGGGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILV ELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFS RYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGI DFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQN TPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK BE4max-REV1-eUNG, SEQ ID NO: 202 MKRTADGSEFESPKKKRKVSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCL LYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCG ECSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYC WRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIAL QSCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKY SIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATR LKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFG NIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNS DVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGL FGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKN LSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQ SKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIP HQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKS EETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTK VKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGV EDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLF DDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHD DSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKP ENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDN VPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETR QITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHH AHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYS NIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKT EVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKS KKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRM LASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEII EQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDT TIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSRRGGWRKRAENDG WETWGGYMAAKVQKLEEQFRSDAAMQKDGTSSTIFSGVAIYVNGYTDPSAEELRK LMMLHGGQYHVYYSRSKTTHIIATNLPNAKIKELKGEKVIRPEWIVESIKAGRLLSYIP YQLYTKQSSVQKGLSFNPVCRPEDPLPGPSNIAKQLNNRVNHIVKKIETENEVKVNG MNSWNEEDENNDFSFVDLEQTSPGRKQNGIPHPRGSTAIFNGHTPSSNGALKTQD CLVPMVNSVASRLSPAFSQEEDKAEKSSTDFRDCTLQQLQQSTRNTDALRNPHRT NSFSLSPLHSNTKINGAHHSTVQGPSSTKSTSSVSTFSKAAPSVPSKPSDCNFISNF YSHSRLHHISMWKCELTEFVNTLQRQSNGIFPGREKLKKMKTGRSALVVTDTGDMS VLNSPRHQSCIMHVDMDCFFVSVGIRNRPDLKGKPVAVTSNRGTGRAPLRPGANP QLEWQYYQNKILKGKAADIPDSSLWENPDSAQANGIDSVLSRAEIASCSYEARQLGI KNGMFFGHAKQLCPNLQAVPYDFHAYKEVAQTLYETLASYTHNIEAVSCDEALVDIT EILAETKLTPDEFANAVRMEIKDQTKCAASVGIGSNILLARMATRKAKPDGQYHLKPE EVDDFIRGQLVTNLPGVGHSMESKLASLGIKTCGDLQYMTMAKLQKEFGPKTGQML YRFCRGLDDRPVRTEKERKSVSAEINYGIRFTQPKEAEAFLLSLSEEIQRRLEATGM KGKRLTLKIMVRKPGAPVETAKFGGHGICDNIARTVTLDQATDNAKIIGKAMLNMFH TMKLNISDMRGVGIHVNQLVPTNLNPSTCPSRPSVQSSHFPSGSYSVRDVFQVQKA KKSTEEEHKEVFRAAVDLEISSASRTCTFLPPFPAHLPTSPDTNKAESSGKWNGLHT PVSVQSRLNLSIEVPSPSQLDQSVLEALPPDLREQVEQVCAVQQAESHGDKKKEPV NGCNTGILPQPVGTVLLQIPEPQESNSDAGINLIALPAFSQVDPEVFAALPAELQREL KAAYDQRQRQGENSTHQQSASASVPKNPLLHLKAAVKEKKRNKKKKTIGSPKRIQS PLNNKLLNSPAKTLPGACGSPQKLIDGFLKHEGPPAEKPLEELSASTSGVPGLSSLQ SDPAGCVRPPAPNLAGAVEFNDVKTLLREWITTISDPMEEDILQVVKYCTDLIEEKDL EKLDLVIKYMKRLMQQSVESVWNMAFDFILDNVQVVLQQTYGSTLKVTSGGSGGS GGSANELTWHDVLAEEKQQPYFLNTLQTVASERQSGVTIYPPQKDVFNAFRFTELG DVKVVILGQDPYHGPGQAHGLAFSVRPGIAIPPSLLNMYKELENTIPGFTRPNHGYL ESWARQGVLLLNTVLTVRAGQAHSHASLGWETFTDKVISLINQHREGVVFLLWGSH AQKKGAIIDKQRHHVLKAPHPSPLSAHRGFFGCNHFVLANQWLEQRGETPIDWMP VLPAESESGGSKRTADGSEFEPKKKRKVGGGGSGATNFSLLKQAGDVEENPGPM VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPW PTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVK FEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIE DGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAG ITLGMDELYK BE4max-REV1-hUNG, SEQ ID NO: 203 MKRTADGSEFESPKKKRKVSSETGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCL LYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCG ECSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYC WRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIAL QSCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKY SIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATR LKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFG NIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNS DVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGL FGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKN LSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQ SKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIP HQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKS EETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTK VKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGV EDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLF DDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHD DSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKP ENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYL YYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDN VPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETR QITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHH AHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYS NIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKT EVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEKGKS KKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRM LASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEI EQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDT TIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSRRGGWRKRAENDG WETWGGYMAAKVQKLEEQFRSDAAMQKDGTSSTIFSGVAIYVNGYTDPSAEELRK LMMLHGGQYHVYYSRSKTTHIIATNLPNAKIKELKGEKVIRPEWIVESIKAGRLLSYIP YQLYTKQSSVQKGLSFNPVCRPEDPLPGPSNIAKQLNNRVNHIVKKIETENEVKVNG MNSWNEEDENNDFSFVDLEQTSPGRKQNGIPHPRGSTAIFNGHTPSSNGALKTQD CLVPMVNSVASRLSPAFSQEEDKAEKSSTDFRDCTLQQLQQSTRNTDALRNPHRT NSFSLSPLHSNTKINGAHHSTVQGPSSTKSTSSVSTFSKAAPSVPSKPSDCNFISNF YSHSRLHHISMWKCELTEFVNTLQRQSNGIFPGREKLKKMKTGRSALVVTDTGDMS VLNSPRHQSCIMHVDMDCFFVSVGIRNRPDLKGKPVAVTSNRGTGRAPLRPGANP QLEWQYYQNKILKGKAADIPDSSLWENPDSAQANGIDSVLSRAEIASCSYEARQLGI KNGMFFGHAKQLCPNLQAVPYDFHAYKEVAQTLYETLASYTHNIEAVSCDEALVDIT EILAETKLTPDEFANAVRMEIKDQTKCAASVGIGSNILLARMATRKAKPDGQYHLKPE EVDDFIRGQLVTNLPGVGHSMESKLASLGIKTCGDLQYMTMAKLQKEFGPKTGQML YRFCRGLDDRPVRTEKERKSVSAEINYGIRFTQPKEAEAFLLSLSEEIQRRLEATGM KGKRLTLKIMVRKPGAPVETAKFGGHGICDNIARTVTLDQATDNAKIIGKAMLNMFH TMKLNISDMRGVGIHVNQLVPTNLNPSTCPSRPSVQSSHFPSGSYSVRDVFQVQKA KKSTEEEHKEVFRAAVDLEISSASRTCTFLPPFPAHLPTSPDTNKAESSGKWNGLHT PVSVQSRLNLSIEVPSPSQLDQSVLEALPPDLREQVEQVCAVQQAESHGDKKKEPV NGCNTGILPQPVGTVLLQIPEPQESNSDAGINLIALPAFSQVDPEVFAALPAELQREL KAAYDQRQRQGENSTHQQSASASVPKNPLLHLKAAVKEKKRNKKKKTIGSPKRIQS PLNNKLLNSPAKTLPGACGSPQKLIDGFLKHEGPPAEKPLEELSASTSGVPGLSSLQ SDPAGCVRPPAPNLAGAVEFNDVKTLLREWITTISDPMEEDILQVVKYCTDLIEEKDL EKLDLVIKYMKRLMQQSVESVWNMAFDFILDNVQVVLQQTYGSTLKVTSGGSGGS GGSIGQKTLYSFFSPSPARKRHAPSPEPAVQGTGVAGVPEESGDAAAIPAKKAPAG QEEPGTPPSSPLSAEQLDRIQRNKAAALLRLAARNVPVGFGESWKKHLSGEFGKPY FIKLMGFVAEERKHYTVYPPPHQVFTWTQMCDIKDVKVVILGQDPYHGPNQAHGLC FSVQRPVPPPPSLENIYKELSTDIEDFVHPGHGDLSGWAKQGVLLLNAVLTVRAHQ ANSHKERGWEQFTDAVVSWLNQNSNGLVFLLWGSYAQKKGSAIDRKRHHVLQTA HPSPLSVYRGFFGCRHFSKTNELLQKSGKKPIDWKELSGGSKRTADGSEFEPKKK RKVGGGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHK FSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHD FFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGH KLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLP DNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK evoFERNY-APOBEC1, SEQ ID NO: 204 MSFERNYDPRELRKETYLLYEIKWGKSGKLWRHWCQNNRTQHAEVYFLENIFNAR RFNPSTHCSITWYLSWSPCAECSQKIVDFLKEHPNVNLEIYVARLYYPENERNRQG LRDLVNSGVTIRIMDLPDYNYCWKTFVSDQGGDEDYWPGHFAPWIKQYSLKL evoAPOBEC1, SEQ ID NO: 205 MSSKTGPVAVDPTLRRRIEPHEFEVFFDPRELRKETCLLYEINWGGRHSIWRHTSQ NTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGECSRAITEFLSRYPNVTLFI YIARLYHLANPRNRQGLRDLISSGVTIQIMTEQESGYCWHNFVNYSPSNESHWPRY PHLWVRLYVLELYCIILGLPPCLNILRRKQSQLTSFTIALQSCHYQRLPPH1LWATGLK BE4max(R33A) w/o UGI = miniCGBE1, SEQ ID NO: 206 MKRTADGSEFESPKKKRKVSSETGPVAVDPTLRRRIEPHEFEVFFDPRELAKETCLL YEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGE CSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCW RNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQ SCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYS IGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRL KRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNL SDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQS KNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSE ETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVE DRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFD DKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDD SLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLY YLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQI TKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIM NFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQ TGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLWAKVEKGKSKKL KSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLAS AGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQI SEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTID RKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKKRKV GGGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSV SGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFK SAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLE YNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNH YLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK nCas9_NG, SEQ ID NO: 207 MKRTADGSEFESPKKKRKVSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKK YSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEAT RLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIF GNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDN SDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNG LFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAK NLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFD QSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSI PHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRK SEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELT KVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISG VEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHL FDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIH DDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHK PENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSD NVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVET RQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYH HAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFY SNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKK TEVQTGGFSKESIRPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGK SKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKR MLASARFLQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDE IIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPRAFKYFD TTIDRKVYRSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKK RKVGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVS GEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKS AMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEY NYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHY LSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKSGGSPKKKRKV CGBE1_NG, SEQ ID NO: 208 MKRTADGSEFESPKKKRKVANELTWHDVLAEEKQQPYFLNTLQTVASERQSGVTIY PPQKDVFNAFRFTELGDVKVVILGQDPYHGPGQAHGLAFSVRPGIAIPPSLLNMYKE LENTIPGFTRPNHGYLESWARQGVLLLNTVLTVRAGQAHSHASLGWETFTDKVISLI NQHREGVVFLLWGSHAQKKGAIIDKQRHHVLKAPHPSPLSAHRGFFGCNHFVLAN QWLEQRGETPIDWMPVLPAESESGGSGGSGGSSSETGPVAVDPTLRRRIEPHEFE VFFDPRELAKETCLLYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTR CSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGV TIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNIL RRKQPQLTFFTIALQSCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSESATPE SSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIG ALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFL VEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFR GHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDL LRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLAR GNSRFAWMTRKSEETITPWNFEEWDKGASAQSFIERMTNFDKNLPNEKVLPKHSL LYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFK KIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDRE MIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDG FANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVD ELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVE NTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTR SDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKA GFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQF YKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEI GKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVL SMPQVNIVKKTEVQTGGFSKESIRPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSV LVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSL FELENGRKRMLASARFLQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFV EQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLG APRAFKYFDTTIDRKVYRSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTAD GSEFEPKKKRKVGGGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVEL DGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRY PDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDF KEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTP IGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK miniCGBE1_NG, SEQ ID NO: 209 MKRTADGSEFESPKKKRKVSSETGPVAVDPTLRRRIEPHEFEVFFDPRELAKETCLL YEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGE CSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCW RNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQ SCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYS IGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRL KRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNL SDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQS KNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSE ETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVE DRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFD DKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDD SLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLY YLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQI TKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIM NFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQ TGGFSKESIRPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKL KSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLAS ARFLQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQIS EFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPRAFKYFDTTIDR KVYRSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKKRKVG GGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVS GEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKS AMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEY NYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHY LSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK nCas9_VRQR, SEQ ID NO: 210 MKRTADGSEFESPKKKRKVSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKK YSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEAT RLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIF GNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDN SDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNG LFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAK NLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFD QSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSI PHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRK SEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELT KVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISG VEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHL FDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIH DDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHK PENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLY LYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSD NVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVET RQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYH HAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFY SNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKK TEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGK SKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKR MLASARELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDE IIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFD TTIDRKQYRSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKK RKVGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVS GEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKS AMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEY NYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHY LSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYKSGGSPKKKRKV CGBE1_VRQR, SEQ ID NO: 211 MKRTADGSEFESPKKKRKVANELTWHDVLAEEKQQPYFLNTLQTVASERQSGVTIY PPQKDVFNAFRFTELGDVKVVILGQDPYHGPGQAHGLAFSVRPGIAIPPSLLNMYKE LENTIPGFTRPNHGYLESWARQGVLLLNTVLTVRAGQAHSHASLGWETFTDKVISLI NQHREGVVFLLWGSHAQKKGAIIDKQRHHVLKAPHPSPLSAHRGFFGCNHFVLAN QWLEQRGETPIDWMPVLPAESESGGSGGSGGSSSETGPVAVDPTLRRRIEPHEFE VFFDPRELAKETCLLYEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTR CSITWFLSWSPCGECSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGV TIQIMTEQESGYCWRNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNIL RRKQPQLTFFTIALQSCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSESATPE SSGGSSGGSDKKYSIGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIG ALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFL VEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFR GHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLE NLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQ IGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVR QQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDL LRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLAR GNSRFAWMTRKSEETITPWNFEEWDKGASAQSFIERMTNFDKNLPNEKVLPKHSL LYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFK KIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDRE MIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDG FANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVD ELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVE NTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTR SDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKA GFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQF YKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEI GKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVL SMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVL VVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSL FELENGRKRMLASARELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFV EQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLG APAAFKYFDTTIDRKQYRSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTAD GSEFEPKKKRKVGGGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVEL DGDVNGHKFSVSGEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRY PDHMKQHDFFKSAMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDF KEDGNILGHKLEYNYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTP IGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK miniCGBE1_VRQR, SEQ ID NO: 212 MKRTADGSEFESPKKKRKVSSETGPVAVDPTLRRRIEPHEFEVFFDPRELAKETCLL YEINWGGRHSIWRHTSQNTNKHVEVNFIEKFTTERYFCPNTRCSITWFLSWSPCGE CSRAITEFLSRYPHVTLFIYIARLYHHADPRNRQGLRDLISSGVTIQIMTEQESGYCW RNFVNYSPSNEAHWPRYPHLWVRLYVLELYCIILGLPPCLNILRRKQPQLTFFTIALQ SCHYQRLPPHILWATGLKSGGSSGGSSGSETPGTSESATPESSGGSSGGSDKKYS IGLAIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGETAEATRL KRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHERHPIFGN IVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDLNPDNSD VDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGEKKNGLF GNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADLFLAAKNL SDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYKEIFFDQS KNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFDNGSIPH QIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAWMTRKSE ETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVYNELTKV KYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSVEISGVE DRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKTYAHLFD DKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFMQLIHDD SLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMGRHKPE NIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNEKLYLY YLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGKSDNV PSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLVETRQI TKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINNYHHAH DAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYFFYSNIM NFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIVKKTEVQ TGGFSKESILPKRNSDKLIARKKDWDPKKYGGFVSPTVAYSVLVVAKVEKGKSKKLK SVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGRKRMLASA RELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYLDEIIEQIS EFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKYFDTTIDR KQYRSTKEVLDATLIHQSITGLYETRIDLSQLGGDSGGSKRTADGSEFEPKKKRKVG GGGSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVS GEGEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKS AMPEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEY NYNSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHY LSTQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK nCas9(H840A) for PE2, SEQ ID NO: 213 MKRTADGSEFESPKKKRKVDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNT DRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDS FFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIY LALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILS ARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTY DDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQ DLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEE LLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRI PYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLP NEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVT VKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDI VLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSG KTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIK KGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKEL GSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLK DDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAE RGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKL VSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVR KMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGR DFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGF DSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKK DLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPE DNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENI IHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD SGGSSGGSSGSETPGTSESATPESSGGSSGGSSSGGSKRTADGSEFEPKKKRKV GSGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGE GEGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAM PEGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNY NSHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLS TQSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK PE2, SEQ ID NO: 214 MKRTADGSEFESPKKKRKVDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNT DRHSIKKNLIGALLFDSGETAEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDS FFHRLEESFLVEEDKKHERHPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIY LALAHMIKFRGHFLIEGDLNPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILS ARLSKSRRLENLIAQLPGEKKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTY DDDLDNLLAQIGDQYADLFLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQ DLTLLKALVRQQLPEKYKEIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEE LLVKLNREDLLRKQRTFDNGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRI PYYVGPLARGNSRFAWMTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLP NEKVLPKHSLLYEYFTVYNELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVT VKQLKEDYFKKIECFDSVEISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDI VLTLTLFEDREMIEERLKTYAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSG KTILDFLKSDGFANRNFMQLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIK KGILQTVKVVDELVKVMGRHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKEL GSQILKEHPVENTQLQNEKLYLYYLQNGRDMYVDQELDINRLSDYDVDAIVPQSFLK DDSIDNKVLTRSDKNRGKSDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAE RGGLSELDKAGFIKRQLVETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKL VSDFRKDFQFYKVREINNYHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVR KMIAKSEQEIGKATAKYFFYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGR DFATVRKVLSMPQVNIVKKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGF DSPTVAYSVLVVAKVEKGKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKK DLIIKLPKYSLFELENGRKRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPE DNEQKQLFVEQHKHYLDEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENI IHLFTLTNLGAPAAFKYFDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGD SGGSSGGSSGSETPGTSESATPESSGGSSGGSSTLNIEDEYRLHETSKEPDVSLG STWLSDFPQAWAETGGMGLAVRQAPLIIPLKATSTPVSIKQYPMSQEARLGIKPHIQ RLLDQGILVPCQSPWNTPLLPVKKPGTNDYRPVQDLREVNKRVEDIHPTVPNPYNL LSGLPPSHQWYTVLDLKDAFFCLRLHPTSQPLFAFEWRDPEMGISGQLTWTRLPQ GFKNSPTLFNEALHRDLADFRIQHPDLILLQYVDDLLLAATSELDCQQGTRALLQTLG NLGYRASAKKAQICQKQVKYLGYLLKEGQRWLTEARKETVMGQPTPKTPRQLREF LGKAGFCRLFIPGFAEMAAPLYPLTKPGTLFNWGPDQQKAYQEIKQALLTAPALGLP DLTKPFELFVDEKQGYAKGVLTQKLGPWRRPVAYLSKKLDPVAAGWPPCLRMVAAI AVLTKDAGKLTMGQPLVILAPHAVEALVKQPPDRWLSNARMTHYQALLLDTDRVQF GPVVALNPATLLPLPEEGLQHNCLDILAEAHGTRPDLTDQPLPDADHTWYTDGSSL LQEGQRKAGAAVTTETEVIWAKALPAGTSAQRAELIALTQALKMAEGKKLNVYTDS RYAFATAHIHGEIYRRRGWLTSEGKEIKNKDEILALLKALFLPKRLSIIHCPGHQKGHS AEARGNRMADQAARKAAITETPDTSTLLIENSSPSGGSKRTADGSEFEPKKKRKVG SGATNFSLLKQAGDVEENPGPMVSKGEELFTGVVPILVELDGDVNGHKFSVSGEG EGDATYGKLTLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMP EGYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYN SHNVYIMADKQKNGIKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLST QSALSKDPNEKRDHMVLLEFVTAAGITLGMDELYK Wild type Cas9, SEQ ID NO: 215 MDKKYSIGLDIGTNSVGWAVITDEYKVPSKKFKVLGNTDRHSIKKNLIGALLFDSGET AEATRLKRTARRRYTRRKNRICYLQEIFSNEMAKVDDSFFHRLEESFLVEEDKKHER HPIFGNIVDEVAYHEKYPTIYHLRKKLVDSTDKADLRLIYLALAHMIKFRGHFLIEGDL NPDNSDVDKLFIQLVQTYNQLFEENPINASGVDAKAILSARLSKSRRLENLIAQLPGE KKNGLFGNLIALSLGLTPNFKSNFDLAEDAKLQLSKDTYDDDLDNLLAQIGDQYADL FLAAKNLSDAILLSDILRVNTEITKAPLSASMIKRYDEHHQDLTLLKALVRQQLPEKYK EIFFDQSKNGYAGYIDGGASQEEFYKFIKPILEKMDGTEELLVKLNREDLLRKQRTFD NGSIPHQIHLGELHAILRRQEDFYPFLKDNREKIEKILTFRIPYYVGPLARGNSRFAW MTRKSEETITPWNFEEVVDKGASAQSFIERMTNFDKNLPNEKVLPKHSLLYEYFTVY NELTKVKYVTEGMRKPAFLSGEQKKAIVDLLFKTNRKVTVKQLKEDYFKKIECFDSV EISGVEDRFNASLGTYHDLLKIIKDKDFLDNEENEDILEDIVLTLTLFEDREMIEERLKT YAHLFDDKVMKQLKRRRYTGWGRLSRKLINGIRDKQSGKTILDFLKSDGFANRNFM QLIHDDSLTFKEDIQKAQVSGQGDSLHEHIANLAGSPAIKKGILQTVKVVDELVKVMG RHKPENIVIEMARENQTTQKGQKNSRERMKRIEEGIKELGSQILKEHPVENTQLQNE KLYLYYLQNGRDMYVDQELDINRLSDYDVDHIVPQSFLKDDSIDNKVLTRSDKNRGK SDNVPSEEVVKKMKNYWRQLLNAKLITQRKFDNLTKAERGGLSELDKAGFIKRQLV ETRQITKHVAQILDSRMNTKYDENDKLIREVKVITLKSKLVSDFRKDFQFYKVREINN YHHAHDAYLNAVVGTALIKKYPKLESEFVYGDYKVYDVRKMIAKSEQEIGKATAKYF FYSNIMNFFKTEITLANGEIRKRPLIETNGETGEIVWDKGRDFATVRKVLSMPQVNIV KKTEVQTGGFSKESILPKRNSDKLIARKKDWDPKKYGGFDSPTVAYSVLVVAKVEK GKSKKLKSVKELLGITIMERSSFEKNPIDFLEAKGYKEVKKDLIIKLPKYSLFELENGR KRMLASAGELQKGNELALPSKYVNFLYLASHYEKLKGSPEDNEQKQLFVEQHKHYL DEIIEQISEFSKRVILADANLDKVLSAYNKHRDKPIREQAENIIHLFTLTNLGAPAAFKY FDTTIDRKRYTSTKEVLDATLIHQSITGLYETRIDLSQLGGDGSPKKKRKVSSDYKDH DGDYKDHDIDYKDDDDK SEQ ID NO: 318 >sp|P97931|UNG_MOUSE Uracil-DNA glycosylase OS = Mus musculus OX = 10090 GN = Ung PE = 1 SV = 3 MIGQKTLYSFFSPTPTGKRTTRSPEPVPGSGVAAEIGGDAVASPAKKARVEQNEQG SPLSAEQLVRIQRNKAAALLRLAARNVPAGFGESWKQQLCGEFGKPYFVKLMGFV AEERNHHKVYPPPEQVFTWTQMCDIRDVKVVILGQDPYHGPNQAHGLCFSVQRPV PPPPSLENIFKELSTDIDGFVHPGHGDLSGWARQGVLLLNAVLTVRAHQANSHKER GWEQFTDAVVSWLNQNLSGLVFLLWGSYAQKKGSVIDRKRHHVLQTAHPSPLSVH RGFLGCRHFSKANELLQKSGKKPINWKEL SEQ ID NO: 319 >tr|Q5BK44|Q5BK44_RAT Uracil-DNA glycosylase OS = Rattus norvegicus OX = 10116GN = Ung PE = 2 SV = 1 MGILGPRPLKLARSLRAPRGARLRSLTPDPDSWQASPAKKARVEQDEPATPPSSPL SAEQLVRIQRNKAAALLRLAARNVPAGLGESWKQQLCGEFGKPYFVKLMGFVAEE RKHHKVYPPPEQVFTWTQMCDIRDVKVVILGQDPYHGPNQAHGLCFSVQRPVPPP PSLENIFKELSTDIDGFVHPGHGDLSGWARQGVLLLNAVLTVRAHQANSHKERGWE QFTDAVVSWLNQNLNGLVFLLWGSYAQKKGSAIDRKRHHVLQTAHPSPLSVYRGF FGCRHFSKANELLQRSGKKPISWKEL SEQ ID NO: 320 >sp|P12887|UNG_YEAST Uracil-DNA glycosylase OS = Saccharomyces cerevisiae (strain ATCC 204508/S288c) OX = 559292 GN = UNG1 PE = 1 SV = 1 MWCMRRLPTNSVMTVARKRKQTTIEDFFGTKKSTNEAPNKKGKSGATFMTITNGA AIKTETKAVAKEANTDKYPANSNAKDVYSKNLSSNLRTLLSLELETIDDSWFPHLMD EFKKPYFVKLKQFVTKEQADHTVFPPAKDIYSWTRLTPFNKVKVVIIGQDPYHNFNQ AHGLAFSVKPPTPAPPSLKNIYKELKQEYPDFVEDNKVGDLTHWASQGVLLLNTSLT VRAHNANSHSKHGWETFTKRVVQLLIQDREADGKSLVFLLWGNNAIKLVESLLGST SVGSGSKYPNIMVMKSVHPSPLSASRGFFGTNHFKMINDWLYNTRGEKMIDWSVV PGTSLREVQEANARLESESKDP SEQ ID NO: 321 >sp|Q9U221|UNG_CAEEL Uracil-DNA glycosylase OS = Caenorhabditis elegans OX = 6239 GN = ung-1 PE = 1 SV = 1 MSKTVRIPDMFLKASAASKRKSASNTENIPEKVPAGNENQEVKKMKLQAPEPTEILL KSLLTGESWSKLLEEEFKKGYISKIEKFLNSEVNKGKQVFPPPTQIFTTFNLLPFDEIS VVIIGQDPYHDDNQAHGLSFSVQKGVKPPPSLKNIYKELESDIEGFKRPDHGNLLGW TRQGVFMLNATLTVRAHEANSHAKIGWQTFTDTVIRIISRQSEKPIVFLLWGGFAHK KEELIDTKKHVVIKTAHPSPLSARKWWGCKCFSKCNTELENSGRNPINWADL SEQ ID NO: 322 >sp|Q9LIH6|UNG_ARATH Uracil-DNA glycosylase, mitochondrial OS = Arabidopsis thaliana OX = 3702 GN = UNG PE = 1 SV = 1 MASSTPKTLMDFFQPAKRLKASPSSSSFPAVSVAGGSRDLGSVANSPPRVTVTTSV ADDSSGLTPEQIARAEFNKFVAKSKRNLAVCSERVTKAKSEGNCYVPLSELLVEES WLKALPGEFHKPYAKSLSDFLEREIITDSKSPLIYPPQHLIFNALNTTPFDRVKTVIIGQ DPYHGPGQAMGLSFSVPEGEKLPSSLLNIFKELHKDVGCSIPRHGNLQKWAVQGVL LLNAVLTVRSKQPNSHAKKGWEQFTDAVIQSISQQKEGVVFLLWGRYAQEKSKLID ATKHHILTAAHPSGLSANRGFFDCRHFSRANQLLEEMGIPPIDWQL SEQ ID NO: 323 >tr|Q7ZVD1|Q7ZVD1_DANRE Uracil-DNA glycosylase OS = Danio rerio OX = 7955 GN = unga PE = 2 SV = 1 MIGQKSIKSFFSPASKKRNLDEIKTGETRDDVKKQKLESGNEAPLSPEQLERIAKNK KAA LERLQSAAPDGIGESWLKALSAEFGKSYFKSLMSFVGEERKKHTIYPPPHAVFTWT QTCDIKDVKVVILGQDPYHGPNQAHGLCFSVQRPVPPPPSLVNIFKELASDIEGFVQ PDHGDLTGWANQGVLLLNAVLTVRAHQANSHKDKGWETFTDAVVHWLSSNMQGL VFILWGSYAQKKGAAINKKQHHVLQAVHPSPLSAHRGFFGCKHFSKANELLKKSGK KPIDWKAL SEQ ID NO: 324 >tr|G1SJ42|G1SJ42_RABIT Uracil-DNA glycosylase OS = Oryctolagus cuniculus OX = 9986 GN = UNG PE = 3 SV = 1 MIGQKTLYSFFSPSPAGKRHTRSPEPAAPGTGVAAATEESRDAEASPAKKARAGKD EPGTPPSSPLSPEQLVRIQRNKAAALLRLAARNVPVGFGESWKKHLCGEFGKPYFI KLMGFVAEERKHHTVYPPPHQVFTWTQMCDIRDVKVVILGQDPYHGPSQAHGLCF SVQRPVPPPPSLENIYKELSTDIEGFVHPGHGDLSGWAKQGVLLLNAVLTVRAHQP TSHKDRGWEQFTDAVVSWLNHNSSGLVFLLWGSYAQRKGSAIDRKRHHVLQTAH PSPLSVYRGFFGCRHFSKTNELLRKSGKKPIDWTKL SEQ ID NO: 325 >tr|A0A452THE0|A0A452THE0_URSMA Uracil-DNA glycosylase OS = Ursus maritimus OX = 29073 GN = UNG PE = 3 SV = 1 MARIQNLNSNSYTGSHARRTLTENKNCDNERALGVWGKGAGSLRLPVHEPRSPEP CKHRGPPKKARAVQEDPGTPPSSPLSPEQLVRIQRNKAAALLRLAARNVPVGFGES WKKPLSAEFGKPYFIKLMGFVAEERKHYTVYPPPHQVFTWTQMCDIRQVKVVILGQ DPYHGPNQAHGLCFSVQRPVPPPPSLENIYKELSTDIDGFVHPGHGDLSGWAKQG VLLLNAVLTVRAHQANSHKERGWEQFTDAVVSWLNQNSSGLVFLLWGSYAQKKG SAIDRKRHHVLQTAHPSPLSVYRGFFGCRHFSKTNELLRKSGKEPINWKDL SEQ ID NO: 326 >tr|A0A2K6MB33|A0A2K6MB33_RHIBE Uracil-DNA glycosylase OS = Rhinopithecus bieti OX = 61621 GN = UNG PE = 3 SV = 1 MIGQKTLYSFFSPSPARKRRAPSPEPAVLGTGVAAVPEENGDAAANPAKKAPAAQE ESGTPSSSPLSAEQLDRIQRNKAAALLRLAARNVPVGFGESWKKHLSGEFGKPYFI KLMGFVAEERKHYTVYPPPHQVFTWTQMCDIRDVKVVILGQDPYHGPNQAHGLCF SVQRPVPPPPSLENIYKELSTDIEDFVHPGHGDLSGWAKQGVLLLNAVLTVRAHQA NSHKERGWEQFTDAVVSWLNQNSNGLVFLLWGSYAQKKGSAIDRKRHHVLQTAH PSPLSVYRGFFGCRHFSKTNELLQKSGKXVKWEFRGLTAFRAGSPEHRFTHIFINS KPVISIVLQILN SEQ ID NO: 327 >tr|A0A4X2KC02|A0A4X2KC02_VOMUR Uracil-DNA glycosylase OS = Vombatus ursinus OX = 29139 GN = UNG PE = 3 SV = 1 MIGQKTLHSFFSPSAPKKRRSCTETPADPGTEAVVQSEDASVSPVRKRRPEDEPRA PSSPLSPEQLDRIQRNKAAALLRLASRNVPAGFGESWKRQLSAEFGKPYFIQLMGF VAEERKRHTVYPPPDQVFTWTQLCEIRDVKVVILGQDPYHGPNQAHGLCFSVQRP VPPPPSLENIYKELSTDIEGFAPPGHGDLSGWARQGVLLLNAVLTVRAHQANSHKE RGWEQFTDAVVSWLNENLDGLVFMLWGSYAQKKGLSINRKRHHVLQTAHPSPLSV HRGFLGCRHFSKTNELLKKSGKKPIDWKAL SEQ ID NO: 328 >tr|A0A1X2AUJ0|A0A1X2AUJ0_9MYCO Uracil-DNA glycosylase OS = Mycobacterium riyadhense OX = 486698 GN = ung PE = 3 SV = 1 MTARPLSELVEQGWAAALAPVTEQVAQMGQFLRTEIAAGRRYLPAGSNVLRAFTFP FDEVRVLIVGQDPYPTPGHAVGLSFSVAPDVRPLPRSLANIFDEYTADLGHPQPSC GDLSPWAQRGVLLLNRVLTVRPSNPASHRGKGWEAVTECAIRALAARSKPLVAILW GRDASTLKPMLATGNCVAIESPHPSPLSASRGFFGSRPFSRANELLAGMGGDPVD WRLP SEQ ID NO: 329 >tr|A0A498LRM7|A0A498LRM7_LABRO Uracil-DNA glycosylase OS = Labeo rohita OX = 84645 GN = UNG PE = 3 SV = 1 MQLSEEQLHQIEQNRRAALERLAKRNVPVPVGESWRKKIGTEFTKPYFTKLMSFVT MERKCFTVYPSPEQVFHCTTLCAIEDVKVVILGQDPYHHPGQAHGLAFSVLRPKPP PPSLENIFMELKEDIVGFRHPGHGDLTGWAKQGVLLLNSVLTVRAHQPTSHEGQG WEIFTDAVVLWLSRNLNGLVFLLWGSYAQRKGRVIDRSLEERCQRILQGMEGSLTA RDRVGIQDFVLLDAYTSETAFMDNLRKRFNENLIYTYIGTLLVSVNPYKELGIYTKKQ MDIYMGVNFFELPPHIFALADNVYRTMISETNNHFILISGESGAGKTEASKKVLQFYA VCCPSTRLLDNVRDRLLLSNPVLEAFGNAKTLKNDNSSRFGKYMDIQFDHQGAAVG GHILSYLLEKSRVVHQNHGERNFHIFYQLVEGGEDELLRWLGLERNCQNYRYLIQG ECAKVSSINDKSDWKTVQKALTIIEFSEKDIEHLFAIIASVLHLGNVHFEASAMGYAKL NSNAEVHWLSKLLGIPSNMLQEGLTHRKIEAKAEEVLSPFTAEHAKYARDALAKAIY GRTFSWLVNKINESLANKWEPVPYFNNKIICDLVEEKHRGIISVLDEECLRPGEATDF TFLEKLEEKMSGHPHFVTHKLADQKTRKTLERGDFRLLHYAGEVTYSVVGFLDKNN DLLYRNIKEVMRQSKNSIIQHCFHTIEPDGKKRPETVATQFKSSLAGLTEILMTKEPW YVRCLKPNHCKQPDRFDDVMVRHQVKYLGLMEHLRVRRAGFAYRRRYEVFLKRR CFSLLLTCEHLTNLNAYLCRYKPLCPDTWPHWKGTPAEGVQRLIKHLGYKPDEYKM GRTKIFIRHPRTLFATEDAFEICKHELATRIQAKYKGYRVKGEYQRQREAATKIETCW RGLQARKERERRAWAVKVIKKFIKGFMNRNQPVSMDNSEYLAFVRQSYLTRLQEN LPKSVLDKTTWLTPPPIMQEYSVPVIKYDRNGFRPRFRQLIFTQAAAYLVEEAKIKQR VNYSSLKGVSVSNLSDNFLILHVTCEDTKQKGDLVLQCSYLFEALTKICVVTKNHNLI KVVQGSVRFDIQPGKEGFVDFKSSSESMVYRAKNGHLMVGDFVDRGYYSLETFTY LLALKAKWPDRITLLRGNHESRQITQVYGFYDECQTKYGNANAWRYCTKVFDMLTV AALIDEQILCVHGGLSPDIKTLDQIRTIERNQEIPHKGAFCDLVWSDPEDVDTWAISP RGAGWLFGAKVTNEFVHINNLKLICRAHQLVHEGYKFMFDEKLVTVWSAPNYCYRC GNIASIMVFKDVNTREPKLFRAVPDSERVIPPRTTTPYFL SEQ ID NO: 330 >sp|O74834|UNG_SCHPO Uracil-DNA glycosylase OS = Schizosaccharomyces pombe (strain 972/ATCC 24843) OX = 284812 GN = ung1 PE = 3 SV = 1 MTVLNTTDKRKADDTVNKLDGKLKQPRLDNFFKTNTSSPALKDTQVLDNKENNSVS KFNKEKWAENLTPAQRKLLQLEIDTLESSWFDALKDEFLKPYFLNLKEFLMKEWQS QRVFPPKEDIYSWSHHTPLHKTKVILLGQDPYHNIGQAHGLCFSVRPGIPCPPSLVNI YKAIKIDYPDFVIPKTGYLVPWADQGILMLNASLTVRAHQAASHSGKGWETFTSAVL QVALNRNRKGLVILAWGTPAAKRLQGLPLKAHYVLRSVHPSPLSAHRGFFECHHFK KTNEWLEEQYGPEKCINWSAVSEQKAKIKSSELESSSTE SEQ ID NO: 331 >tr|A0A3B5KG53|A0A3B5KG53_TAKRU Uracil-DNA glycosylase OS = Takifugu rubripes OX = 31033 GN = ung PE = 3 SV = 1 MIGQKTINSFFSPVPKKRICKDLSETEEDAKDHIIQKKRKSPEPEPASPPAAPLSSEQ LERIARNKRAALERLTSAQIPAGIGEGWRDKLSAEFGKPYFKQLTTYVAEERKRRTV YPPADQVFTWTQMCDIRDVKVVILGQDPYHGHNQAHGLCFSVKRPVPPPPSLENM YKELVSDIPGFQHPGHGDLTGWAKQGVLLLNAVLTVRAHNANSHKDKGWETFTDA VVQWLNTNLDGVVFMLWGSYAQKKGAAINRKRHHVLQTVHPSPLSAHRGFFGCA HFSKANELLKKSGKSPVDWKA L SEQ ID NO: 332 >tr|I3M8Q6|I3M8Q6_ICTTR Uracil-DNA glycosylase OS = Ictidomys tridecemlineatus OX = 43179 GN = UNG PE = 3 SV = 1 MIGQKTLYSFFSPSPARKRSVRSPEPADLGTGVVAVAEENGDAADHPTKKARVGQ EEPDTPPSSPLSQEQLVRIQRNKAAALLRLAARNVPVGFGESWRKPLGAEFGKPYF IKLMGFVAEERKRYTVYPPPHQVFTWTQTCDIKDVKVVILGQDPYHGPNQAHGLCF SVQRPVPPPPSLENIYKELSTDIDGFVHPGHGDLSGWAKQGVLLLNAVLTVRAHQA NSHKERGWEEFTDAVVSWLNQNLNGLVFLLWGSYAQKKGIAIDRKRHHVLQTAHP SPLSVYRGFFGCRHFSKANELLQKSGKKPIDWKEL SEQ ID NO: 333 >tr|A0A3P9H4T8|A0A3P9H4T8_ORYLA Uracil-DNA glycosylase OS = Oryzias latipes OX = 8090 GN = UNG PE = 3 SV = 1 MLWLRHRSCDKLVGRFLGTGSVIRNKMMKNWGVIGGIAAAVAAGVYVLWGPITVK KKRKKGMSPGLLNLGNTCFLNALLQGLAACPSFIRWLEKFSGLPSIQSCKDNQLSTT LLQLLKALSSDEPGEDVLDAGCLLDVLRLYRWHISSFEEQDAHELFHVITSSLEEER DRQPKVTHLFDVQFLESFPNQDDKALTCISRAPLHPLPGSWKFQHPFHGRLTSNMS CKRCETQSPVRYDSFESLSLSILLPQWGRPISLDQCLQHFISSETIKEVECENCTKLQ QHSSINGQLLESQRTTFVKQLRLGKLPQCLCIHLQRLMWSNEGSPIKRQEHVQFSE YLSMDRYKHDSSTPRTQRVRCAPKTIKAESFDSIEKSMANGTEHHNNNKPFLNGTC SSMFLGSGVKNPFGFTHHDNSSAEYLFQLVAVLVHLGDMHSGHFVTYRRSPSSSR SSSNFSSQWLWVSDDSKKLKIAAVDPEPQSSPLSPEQLDKIARNKKAALEKLASGLT PQGFSESWRGELLSEFSKPYFKDLTKFVSDERKRGTVYPPAEQIFTWTQMCDIRDV KVVILGQDPYHGPGQAHGLCFSVKRPVSPPPSLENMYKELVSDIEGFKHPGHGDLT GWAQQGVLLLNAVLTVRAHQANSHKDKGWEVFTDAVVQWLSNNLQGLVFLLWGS YAQKKGSAINRKHHHVLQAVHPSPLSAHRGFFGCKHFSKANELLKKSGKSPIDWKA L SEQ ID NO: 334 >tr|A0A4W4HK79|A0A4W4HK79_ELEEL Uracil-DNA glycosylase OS = Electrophorus electricus OX = 8005 GN = ung PE = 3 SV = 1 MIGQKSIKSFFSPTSKKRDTDEQTRSEDICNVKKFKTNTSAVLPSPSLSPELLEKIAK NKKAAQERLAARSAPEGIGKSWQRALGAEFGKTYFKSLMSFVAEERQKQTIYPPPH QVFTWTRMCEIEDVKVVVLGQDPYHGPNQAHGLCFSVQRPVPPPPSLVNMYKELE ADIEGFRHPGHGDLTGWAKQGVLLLNAVLTVRAHQANSHKDKGWEILTDAVVNWL SANLEGLVFMLWGAYAQKKGAAIDRKRHHVLQAVHPSPLSAHRGFFGCKHFSKTN ELLKKSGKKPIDWKAL SEQ ID NO: 335 >tr|A0A5G3K4Q6|A0A5G3K4Q6_XENTR Uracil-DNA glycosylase OS = Xenopus tropicalis OX = 8364 GN = aoc3 PE = 3 SV = 1 MSHPICRPNMSVMFWLLPFPKLPVLSESWRQTSVVCSIRTKQRIGAGVIIPGFSRGA MIGQRTINSFFGPAAKKRAAPEALGEEGPYKGEITPVKKSRQSGENEIPPAVSPPLS PEQLERIQRNKAAALQKLAARHVPEGLGQSWKQALLAEFAKPYFVKLSNFVAEERK KYTVYPPPEEVFTWTQMVDIKDVKVVILGQDPYHGPNQAHGLCFSVKKPVPPPPSL VNMYKELETDIEGFSHPGHGDLTGWAKQGVLLLNAVLTVRAHNANSHKDCGWEQF TDSVVSWLNKNMDGLVFMLWGAYAQKKGSNIDRKRHLVLQTVHPSPLSAHRGFFG CCHFSKTNAYLQGLGKKPIDWKAL SEQ ID NO: 336 >tr|A0A0F0TTY1|A0A0F0TTY1_ENTCL Uracil-DNA glycosylase OS = Enterobacter cloacae subsp. cloacae OX = 336306 GN = ung PE = 3 SV = 1 MTTPLTWHDVLAEEKQQPYFINTLSTVAAERQSGQTIYPPQKDVFNAFRYTELSDV KWILGQDPYHGPGQAHGLAFSVRPGVAIPPSLLNMYKELEGTIPGFTRPNHGYLES WARQGVLLLNTVLTVRAGQAHSHASLGWETFTDKVISLINEHREGVVFLLWGSHAQ KKGAIIDRQRHHVLKAPHPSPLSAHRGFFGCNHFVLANEWLEKRGETPIDWMPVLP AESE SEQ ID NO: 337 >tr|A0A1V4IJH4|A0A1V4IJH4_9CLOT Uracil-DNA glycosylase OS = Clostridium oryzae OX = 1450648 GN = ung PE = 3 SV = 1 MTVNIKNDWLELLEDQFEMDYYKDLRHFLISEYKTRTIYPDMYDIFNALNYTAYKDVK VVILGQDPYHGPNQAHGLSFSVKPGVPAPPSLINIYKELKDDLGCYIPNNGYLKKWT DEGVLLLNTALTVRAGEANSHRNKGWEIFTDAIISLLNKREKSIVFILWGSNAISKEKLI TNKAHYIIKSPHPSPLSAHRGFFGSKPFSKANNFLKSIGEKPIDWQIENI SEQ ID NO: 338 >tr|A0A1C3ZIJ7|A0A1C3ZIJ7_9LACO Uracil-DNA glycosylase OS = Lactobacillus apis OX = 303541 GN = ung PE = 3 SV = 1 MKKFIGNDWDEVLAPVFESNEYHALHEFLKKEYQTKRIFPDMYHIFTAFKLTPFAKT KWILGQDPYHNPGQATGMSFAVMPGVKLPPSLQNIYKELYDDVGCVPVQHGYLK KWADQGVLLLNAVLTVPYGHANGHQGKGWEQVTDAAIKALSDRGQVVFILWGKYA QNKIALIDQEKNYVIKSAHPSPFSADRGFFGSRPFSRCNEALKKFGEAPIDWQLPQQ VTESDLA SEQ ID NO: 339 >tr|A0A519N079|A0A519N079_FLASP Uracil-DNA glycosylase OS = Flavobacterium sp. OX = 239 GN = ung PE = 3 SV = 1 MKIEESWKKELQSEFEKPYFKELREFISREFDAENGKTCYPPESQIFSAFDHCPFDE VKVVIIGQDPYHGPGQANGLCFSVADGIPIRPSLRNIFVEIKNDLGKPIPATGNLERW ANQGVLLLNATLTVRQGEAGSHQKQGWETFTDAVIQHISDDRQNVVFLLWGAFAQ QKGKNIDKSKHCVLTSGHPSPMSANQGKWFGNKHFSKANEYLKSKGLPEIDW SEQ ID NO: 340 >tr|A0A1H3TI78|A0A1H3TI78_9BURK Uracil-DNA glycosylase OS = Delftia lacustris OX = 558537 GN = ung PE = 3 SV = 1 MALQDDAIAPAQADQLQSADPADWPVAPDWQPLVEDFFAGATGQQLLTFLHQRLE AGAVIFPPQPLRALELTPPDEVRVVILGQDPYHGRGQAEGLSFSVAPGVRMPPSLQ NIFKEMQRDLGVPFPPFPNPGGSLVKWARNGVLLLNTCLTVEEGQAASHSGKGWE LLTDAVIRHIAQGTRPVVFMLWGSHAQSKRAFIPGDRGHLVLTSNHPSPLSALRPPV PFIGNGHFGKARDFRAQHGY SEQ ID NO: 341 >tr|A0A3D4RH89|A0A3D4RH89_9LACT Uracil-DNA glycosylase OS = Lactococcus garvieae OX = 1363 GN = ung PE = 3 SV = 1 MKKTDWSGPLRERLPQEYFSDLVDFINEVYAKGNVYPPEDKIFRAIELTALSDVKVIL VGQDPYPQPGKAQGLSFSYPASFVVNRPDSIVNIRKELQSEGFDKKDSDLTHWAE QGVLLLNAVLTVPEMKSNAHKGKIWEPLTDEIIKIASDDARPKVFLLWGGDARKKAK LIDSSKHLVLESAHPSPLSASRGFFGSQPFSKANAFLEKTGQKGIDWSK SEQ ID NO: 342 >tr|A0A2Z6T8A7|A0A2Z6T8A7_9LACO Uracil-DNA glycosylase OS = Lactobacillus rodentium OX = 947835 GN = ung PE = 3 SV = 1 MKNLIGNDWDEILAPVFQSENYQELHNFLKEEYQTKTIYPDMYHIFTAFKLTPFAKTK VVILGQDPYHNPGQATGMSFSVNPGIALPPSLKNIYKELYDDVGAVPVDHGYLKKW ADQGVLLLNAVLTVPYGKANGHQGKGWEFVTDQAIKRLSERGNVVFILWGRFAQN KIPLIDQNKNFIIKSSHPSPFSADRGFFGSRPFSRCNDALKQFNEAPIDWQLPAKVNR TEIV SEQ ID NO: 343 >sp|Q53HV7|SMUG1_HUMAN Single-strand selective monofunctional uracil DNA glycosylase OS = Homo sapiens OX = 9606 GN = SMUG1 PE = 1 SV = 2 MPQAFLLGSIHEPAGALMEPQPCPGSLAESFLEEELRLNAELSQLQFSEPVGIIYNP VEY AWEPHRNYVTRYCQGPKEVLFLGMNPGPFGMAQTGVPFGEVSMVRDWLGIVGPV LTPPQEHPKRPVLGLECPQSEVSGARFWGFFRNLCGQPEVFFHHCFVHNLCPLLF LAPSGRNLTPAELPAKQREQLLGICDAALCRQVQLLGVRLVVGVGRLAEQRARRAL AGLMPEVQVEGLLHPSPRNPQANKGWEAVAKERLNELGLLPLLLK SEQ ID NO: 344 >sp|Q811Q1|SMUG1_RAT Single-strand selective monofunctional uracil-DNA glycosylase OS = Rattus norvegicus OX = 10116 GN = Smug1 PE = 2 SV = 1 MAVSQTFPPGPAHEPASALMEPCARSLAEGFLEEELRLNAELSQLQFPEPVGVIYN PVDYAWEPHRNYVTRYCQGPKEVLFLGMNPGPFGMAQTGVPFGEVNVVRDWLGI GGSVLSPPQEHPKRPVLGLECPQSEVSGARFWGFFRTLCGQPQVFFRHCFVHNL CPLLFLAPSGRNLTPADLPAKHREQLLSICDAALCRQVQLLGVRLVVGVGRLAEQR ARRALAGLTPEVQVEGLLHPSPRSPQANKGWETAARERLQELGLLPLLTDEGSVRP TP SEQ ID NO: 345 >sp|Q6P5C5|SMUG1_MOUSE Single-strand selective monofunctional uracil DNA glycosylase OS = Mus musculus OX = 10090 GN = Smug1 PE = 1 SV = 1 MAASQTFPLGPTHEPASALMEPLPCTRSLAEGFLEEELRLNAELSQLQFPEPVGVIY NPVDYAWEPHRNYVTRYCQGPKEVLFLGMNPGPFGMAQTGVPFGEVNVVRDWL GVGGPVLTPPQEHPKRPVLGLECPQSEVSGARFWGFFRTLCGQPQVFFRHCFVH NLCPLLFLAPSGRNLTPAELPAKQREQLLSICDAALCRQVQLLGVRLVVGVGRLAEQ RARRALAGLTPEVQVEGLLHPSPRSAQANKGWEAAARERLQELGLLPLLTDEGSA RPT SEQ ID NO: 346 >sp|Q9YGN6|SMUG1_XENLA Single-strand selective monofunctional uracil DNA glycosylase OS = Xenopus laevis OX = 8355 GN = smug1 PE = 1 SV = 1 MAAEACVPAEFSKDEKNGSILSAFCSDIPDITSSTESPADSFLKVELELNLKLSNLVF QD PVQYVYNPLVYAWAPHENYVQTYCKSKKEVLFLGMNPGPFGMAQTGVPFGEVNH VRDWLQIEGPVSKPEVEHPKRRIRGFECPQSEVSGARFWSLFKSLCGQPETFFKH CFVHNHCPLIFMNHSGKNLTPTDLPKAQRDTLLEICDEALCQAVRVLGVKLVIGVGR FSEQRARKALMAEGIDVTVKGIMHPSPRNPQANKGWEGIVRGQLLELGVLSLLTG SEQ ID NO: 347 >sp|Q59l47|SMUG1_BOVIN Single-strand selective monofunctional uracil DNA glycosylase OS = Bos taurus OX = 9913 GN = SMUG1 PE = 2 SV = 1 MAVPQPFPSGPHLQPAGALMEPQPSPRSLAEGFLQEELRLNDELRQLQFSELVGIV YNPVEYAWEPHRSYVTRYCQGPKQVLFLGMNPGPFGMAQTGVPFGEVSVVRDWL GLGGPVRTPPQEHPKRPVLGLECPQSEVSGARFWGFFRNLCGQPEVFFRHCFVH NLCPLLLLAPSGRNITPAELPAKQREQLLGVCDAALCRQVQLLGVRLVVGVGRVAE QRARRALASLMPEVQVEGLLHPSPRSPQANKGWEAVAKERLNELGLLPLLTS

OTHER EMBODIMENTS

It is to be understood that while the invention has been described in conjunction with the detailed description thereof, the foregoing description is intended to illustrate and not limit the scope of the invention, which is defined by the scope of the appended claims. Other aspects, advantages, and modifications are within the scope of the following claims. 

1. A C-to-G transversion base editor (CGBE) comprising a cytidine deaminase, a programmable DNA binding domain, and further comprising one or more nuclear localization sequences (NLS), and optionally one or more human or E. coli or other uracil-n-glycosylases (UNGs) or SMUG1, preferably wherein the CGBE does not comprise a uracil-N-glycosylase inhibitor (UGI).
 2. The CGBE of claim 1, wherein the cytidine deaminase comprises an active cytidine deaminase domain from an engineered rat APOBEC1 (rAPOBEC1) comprising a mutation at residue R33.
 3. (canceled)
 4. The CGBE of claim 1, wherein the rAPOBEC1 further comprises one or more mutations at amino acid positions that correspond to residues P29, K34, E181, and/or L182 of rAPOBEC1 (SEQ ID NO:67) or to W90Y, R126E, R132E, W90Y+R126E (double mutant), R126E+R132E (double mutant), W90Y+R132E (double mutant), W90Y+R126E+R132E (triple mutant).
 5. (canceled)
 6. The CGBE of claim 1, wherein the mutation at amino acid position that correspond to residue R33 is a R33A substitution mutation.
 7. The CGBE of claim 1, wherein the CGBE comprises N- or C-terminal fusions of one or more human or E. coli UNG or SMUG1 or other orthologues of UNG or SMUG1.
 8. The CGBE of claim 7, wherein the one or more UNGs are from E. coli.
 9. The CGBE of claim 1, where the UNG(s) is absent.
 10. The CGBE of claim 1, wherein the rAPOBEC1 comprises a R33A mutation and one or more mutations at positions: P29F, P29T, K34A, E181Q and/or L182A of rAPOBEC1 (SEQ ID NO:67).
 11. The CGBE of claim 10, further comprising one or more mutations in the rAPOBEC1 at residues corresponding to E24, V25; R118, Y120, H121, R126; W224-K229; P168-1186; L173+L180; R15, R16, R17, to K15-17 & A15-17; Deletion E181-L210; P190+P191; Deletion L210-K229 (C-terminal); and/or Deletion S2-L14 (N-terminal) of SEQ ID NO:67.
 12. (canceled)
 13. (canceled)
 14. (canceled)
 15. The CGBE of claim 1, comprising a linker between the cytosine deaminase and/or between the cytosine deaminase or single-chain dimers and the programmable DNA binding domain.
 16. The CGBE of claim 1, wherein the programmable DNA binding domain is selected from the group consisting of a Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) Cas RNA-guided nuclease (RGN), an engineered C2H2 zinc-finger, a transcription activator effector-like effector (TALE), and variants thereof.
 17. The CGBE of claim 1, wherein the CRISPR RGN is a ssDNA nickase or a catalytically inactive CRISPR Cas RNA-guided nuclease, optionally a Cas9 or Cas12a that has ssDNA nickase activity or is catalytically inactive.
 18. A base editing system comprising: (i) an CGBE of claim 1, wherein the programmable DNA binding domain is a CRISPR Cas RGN or a variant thereof; and (ii) at least one guide RNA compatible with the base editor comprising a spacer sequence that directs the base editor to a target sequence, preferably wherein the target sequence comprises a cytosine at position 4-8, 5-7, or position 6 (with 1 being the most PAM-distal position).
 19. An isolated nucleic acid encoding a CGBE of claim
 1. 20. A vector comprising the isolated nucleic acid of claim
 19. 21. An isolated host cell, preferably a mammalian host cell, comprising the nucleic acid of claim
 19. 22. The isolated host cell of claim 21, wherein the isolated host cell expresses a CGBE.
 23. A composition comprising: (i) a CGBE of claim 1, wherein the programmable DNA binding domain is a CRISPR Cas RGN or a variant thereof; (ii) at least one guide RNA compatible with the base editor comprising a spacer sequence that directs the base editor to a target sequence, preferably wherein the target sequence comprises a cytosine at position 4-8, 5-7, or position 6 (with 1 being the most PAM-distal position), and (iii) a pharmaceutically acceptable carrier.
 24. The composition of claim 23, comprising one or more ribonucleoprotein (RNP) complexes.
 25. A method of generating a cytosine-to-guanine and guanine-to-cytosine alteration in a nucleic acid, the method comprising contacting the nucleic acid with the CGBE of claim
 1. 26. (canceled)
 27. (canceled)
 28. (canceled)
 29. (canceled)
 30. (canceled)
 31. (canceled)
 32. (canceled)
 33. (canceled)
 34. (canceled) 